U.S. patent application number 17/569487 was filed with the patent office on 2022-06-30 for anti-sars-cov-2 antibodies and uses thereof.
The applicant listed for this patent is TSB THERAPEUTICS (BEIJING) CO., LTD.. Invention is credited to Bin JU, Lei LIU, Xuanling SHI, Linqi ZHANG, Qi ZHANG, Zheng ZHANG, Qing ZHU.
Application Number | 20220204591 17/569487 |
Document ID | / |
Family ID | |
Filed Date | 2022-06-30 |
United States Patent
Application |
20220204591 |
Kind Code |
A1 |
ZHANG; Zheng ; et
al. |
June 30, 2022 |
ANTI-SARS-COV-2 ANTIBODIES AND USES THEREOF
Abstract
Provided herein are modified anti-SARS-COV-2 antibodies or
antigen binding fragments thereof having extended half life and
optimized immune activities. Disclosed herein is also directed to
pharmaceutical compositions comprising the same and a method for
treating or preventing a disease in human patients that is caused
by or related to the infection of SARS-COV-2.
Inventors: |
ZHANG; Zheng; (Shenzhen,
CN) ; ZHANG; Linqi; (Beijing, CN) ; LIU;
Lei; (Shenzhen, CN) ; ZHANG; Qi; (Beijing,
CN) ; JU; Bin; (Shenzhen, CN) ; SHI;
Xuanling; (Beijing, CN) ; ZHU; Qing; (Beijing,
CN) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
TSB THERAPEUTICS (BEIJING) CO., LTD. |
Beijing |
|
CN |
|
|
Appl. No.: |
17/569487 |
Filed: |
January 5, 2022 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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16953304 |
Nov 19, 2020 |
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17569487 |
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International
Class: |
C07K 16/10 20060101
C07K016/10 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 20, 2020 |
CN |
202010203065.1 |
Mar 21, 2020 |
CN |
PCT/CN2020/080532 |
Apr 10, 2020 |
CN |
PCT/CN2020/084097 |
Apr 14, 2020 |
CN |
PCT/CN2020/084805 |
Aug 12, 2020 |
CN |
PCT/CN2020/108718 |
Claims
1-34. (canceled)
35. An antibody or an antigen-binding fragment thereof having
specific binding affinity to receptor binding domain (RBD) of spike
protein of SARS-CoV-2, wherein said antibody or antigen-binding
domain comprises: a. a heavy chain CDR1 (HCDR1) comprising the
sequence of SEQ ID NO: 1, a heavy chain CDR2 (HCDR2) comprising the
sequence of SEQ ID NO: 2, a heavy chain CDR3 (HCDR3) comprising the
sequence of SEQ ID NO: 3; a light chain CDR1 (LCDR1) comprising the
sequence of SEQ ID NO: 4, a light chain CDR2 (LCDR2) comprising the
sequence of SEQ ID NO: 5, and a light chain CDR3 (LCDR3) comprising
the sequence of SEQ ID NO: 6; b. a HCDR1 comprising the sequence of
SEQ ID NO: 11, a HCDR2 comprising the sequence of SEQ ID NO: 12, a
HCDR3 comprising the sequence of SEQ ID NO: 13, a LCDR1 comprising
the sequence of SEQ ID NO: 14, a LCDR2 comprising the sequence of
SEQ ID NO: 15, and a LCDR3 comprising the sequence of SEQ ID NO:
16; c. a HCDR1 comprising the sequence of SEQ ID NO: 21, a HCDR2
comprising the sequence of SEQ ID NO: 22, a HCDR3 comprising the
sequence of SEQ ID NO: 23, a LCDR1 comprising the sequence of SEQ
ID NO: 24, a LCDR2 comprising the sequence of SEQ ID NO: 25, and a
LCDR3 comprising the sequence of SEQ ID NO: 26; d. a HCDR1
comprising the sequence of SEQ ID NO: 31, a HCDR2 comprising the
sequence of SEQ ID NO: 32, a HCDR3 comprising the sequence of SEQ
ID NO: 33, a LCDR1 comprising the sequence of SEQ ID NO: 34, a
LCDR2 comprising the sequence of SEQ ID NO: 35, and a LCDR3
comprising the sequence of SEQ ID NO: 36; e. a HCDR1 comprising the
sequence of SEQ ID NO: 41, a HCDR2 comprising the sequence of SEQ
ID NO: 42, a HCDR3 comprising the sequence of SEQ ID NO: 43, a
LCDR1 comprising the sequence of SEQ ID NO: 44, a LCDR2 comprising
the sequence of SEQ ID NO: 45, and a LCDR3 comprising the sequence
of SEQ ID NO: 46; f. a HCDR1 comprising the sequence of SEQ ID NO:
51, a HCDR2 comprising the sequence of SEQ ID NO: 52, a HCDR3
comprising the sequence of SEQ ID NO: 53, a LCDR1 comprising the
sequence of SEQ ID NO: 54, a LCDR2 comprising the sequence of SEQ
ID NO: 55, and a LCDR3 comprising the sequence of SEQ ID NO: 56; g.
a HCDR1 comprising the sequence of SEQ ID NO: 65, a HCDR2
comprising the sequence of SEQ ID NO: 66, a HCDR3 comprising the
sequence of SEQ ID NO: 67, a LCDR1 comprising the sequence of SEQ
ID NO: 68, a LCDR2 comprising the sequence of SEQ ID NO: 69, and a
LCDR3 comprising the sequence of SEQ ID NO: 70; h. a HCDR1
comprising the sequence of SEQ ID NO: 75, a HCDR2 comprising the
sequence of SEQ ID NO: 76, a HCDR3 comprising the sequence of SEQ
ID NO: 77, a LCDR1 comprising the sequence of SEQ ID NO: 78, a
LCDR2 comprising the sequence of SEQ ID NO: 79, and a LCDR3
comprising the sequence of SEQ ID NO: 80; i. a HCDR1 comprising the
sequence of SEQ ID NO: 85, a HCDR2 comprising the sequence of SEQ
ID NO: 86, a HCDR3 comprising the sequence of SEQ ID NO: 87, a
LCDR1 comprising the sequence of SEQ ID NO: 88, a LCDR2 comprising
the sequence of SEQ ID NO: 89, and a LCDR3 comprising the sequence
of SEQ ID NO: 90; j. a HCDR1 comprising the sequence of SEQ ID NO:
95, a HCDR2 comprising the sequence of SEQ ID NO: 96, a HCDR3
comprising the sequence of SEQ ID NO: 97, a LCDR1 comprising the
sequence of SEQ ID NO: 98, a LCDR2 comprising the sequence of SEQ
ID NO: 99, and a LCDR3 comprising the sequence of SEQ ID NO: 100;
k. a HCDR1 comprising the sequence of SEQ ID NO: 105, a HCDR2
comprising the sequence of SEQ ID NO: 106, a HCDR3 comprising the
sequence of SEQ ID NO: 107, a LCDR1 comprising the sequence of SEQ
ID NO: 108, a LCDR2 comprising the sequence of SEQ ID NO: 109, and
a LCDR3 comprising the sequence of SEQ ID NO: 110; l. a HCDR1
comprising the sequence of SEQ ID NO: 136, a HCDR2 comprising the
sequence of SEQ ID NO: 137, a HCDR3 comprising the sequence of SEQ
ID NO: 138, a LCDR1 comprising the sequence of SEQ ID NO: 139, a
LCDR2 comprising the sequence of SEQ ID NO: 140, and a LCDR3
comprising the sequence of SEQ ID NO: 141; m. HCDR1 comprising the
sequence of SEQ ID NO: 146, a HCDR2 comprising the sequence of SEQ
ID NO: 147, a HCDR3 comprising the sequence of SEQ ID NO: 148, a
LCDR1 comprising the sequence of SEQ ID NO: 149, a LCDR2 comprising
the sequence of SEQ ID NO: 150, and a LCDR3 comprising the sequence
of SEQ ID NO: 151; n. HCDR1 comprising the sequence of SEQ ID NO:
156, a HCDR2 comprising the sequence of SEQ ID NO: 157, a HCDR3
comprising the sequence of SEQ ID NO: 158, a LCDR1 comprising the
sequence of SEQ ID NO: 159, a LCDR2 comprising the sequence of SEQ
ID NO: 160, and a LCDR3 comprising the sequence of SEQ ID NO: 161;
o. HCDR1 comprising the sequence of SEQ ID NO: 166, a HCDR2
comprising the sequence of SEQ ID NO: 167, a HCDR3 comprising the
sequence of SEQ ID NO: 168, a LCDR1 comprising the sequence of SEQ
ID NO: 169, a LCDR2 comprising the sequence of SEQ ID NO: 170, and
a LCDR3 comprising the sequence of SEQ ID NO: 171; p. HCDR1
comprising the sequence of SEQ ID NO: 176, a HCDR2 comprising the
sequence of SEQ ID NO: 177, a HCDR3 comprising the sequence of SEQ
ID NO: 178, a LCDR1 comprising the sequence of SEQ ID NO: 179, a
LCDR2 comprising the sequence of SEQ ID NO: 180, and a LCDR3
comprising the sequence of SEQ ID NO: 181; q. HCDR1 comprising the
sequence of SEQ ID NO: 186, a HCDR2 comprising the sequence of SEQ
ID NO: 187, a HCDR3 comprising the sequence of SEQ ID NO: 188, a
LCDR1 comprising the sequence of SEQ ID NO: 189, a LCDR2 comprising
the sequence of SEQ ID NO: 190, and a LCDR3 comprising the sequence
of SEQ ID NO: 191; r. HCDR1 comprising the sequence of SEQ ID NO:
196, a HCDR2 comprising the sequence of SEQ ID NO: 197, a HCDR3
comprising the sequence of SEQ ID NO: 198, a LCDR1 comprising the
sequence of SEQ ID NO: 199, a LCDR2 comprising the sequence of SEQ
ID NO: 200, and a LCDR3 comprising the sequence of SEQ ID NO: 201;
s. HCDR1 comprising the sequence of SEQ ID NO: 206, a HCDR2
comprising the sequence of SEQ ID NO: 207, a HCDR3 comprising the
sequence of SEQ ID NO: 208, a LCDR1 comprising the sequence of SEQ
ID NO: 209, a LCDR2 comprising the sequence of SEQ ID NO: 210, and
a LCDR3 comprising the sequence of SEQ ID NO: 211; t. HCDR1
comprising the sequence of SEQ ID NO: 216, a HCDR2 comprising the
sequence of SEQ ID NO: 217, a HCDR3 comprising the sequence of SEQ
ID NO: 218, a LCDR1 comprising the sequence of SEQ ID NO: 219, a
LCDR2 comprising the sequence of SEQ ID NO: 220, and a LCDR3
comprising the sequence of SEQ ID NO: 221; u. HCDR1 comprising the
sequence of SEQ ID NO: 226, a HCDR2 comprising the sequence of SEQ
ID NO: 227, a HCDR3 comprising the sequence of SEQ ID NO: 228, a
LCDR1 comprising the sequence of SEQ ID NO: 229, a LCDR2 comprising
the sequence of SEQ ID NO: 230, and a LCDR3 comprising the sequence
of SEQ ID NO: 231; v. HCDR1 comprising the sequence of SEQ ID NO:
236, a HCDR2 comprising the sequence of SEQ ID NO: 237, a HCDR3
comprising the sequence of SEQ ID NO: 238, a LCDR1 comprising the
sequence of SEQ ID NO: 239, a LCDR2 comprising the sequence of SEQ
ID NO: 240, and a LCDR3 comprising the sequence of SEQ ID NO: 241;
w. HCDR1 comprising the sequence of SEQ ID NO: 246, a HCDR2
comprising the sequence of SEQ ID NO: 247, a HCDR3 comprising the
sequence of SEQ ID NO: 248, a LCDR1 comprising the sequence of SEQ
ID NO: 249, a LCDR2 comprising the sequence of SEQ ID NO: 250, and
a LCDR3 comprising the sequence of SEQ ID NO: 251; x. HCDR1
comprising the sequence of SEQ ID NO: 256, a HCDR2 comprising the
sequence of SEQ ID NO: 257, a HCDR3 comprising the sequence of SEQ
ID NO: 258, a LCDR1 comprising the sequence of SEQ ID NO: 259, a
LCDR2 comprising the sequence of SEQ ID NO: 260, and a LCDR3
comprising the sequence of SEQ ID NO: 261; y. HCDR1 comprising the
sequence of SEQ ID NO: 266, a HCDR2 comprising the sequence of SEQ
ID NO: 267, a HCDR3 comprising the sequence of SEQ ID NO: 268, a
LCDR1 comprising the sequence of SEQ ID NO: 269, a LCDR2 comprising
the sequence of SEQ ID NO: 270, and a LCDR3 comprising the sequence
of SEQ ID NO: 271; z. HCDR1 comprising the sequence of SEQ ID NO:
276, a HCDR2 comprising the sequence of SEQ ID NO: 277, a HCDR3
comprising the sequence of SEQ ID NO: 278, a LCDR1 comprising the
sequence of SEQ ID NO: 279, a LCDR2 comprising the sequence of SEQ
ID NO: 280, a LCDR3 comprising the sequence of SEQ ID NO: 281; aa.
HCDR1 comprising the sequence of SEQ ID NO: 286, a HCDR2 comprising
the sequence of SEQ ID NO: 287, a HCDR3 comprising the sequence of
SEQ ID NO: 288, a LCDR1 comprising the sequence of SEQ ID NO: 289,
a LCDR2 comprising the sequence of SEQ ID NO: 290, a LCDR3
comprising the sequence of SEQ ID NO: 291; bb. HCDR1 comprising the
sequence of SEQ ID NO: 296, a HCDR2 comprising the sequence of SEQ
ID NO: 297, a HCDR3 comprising the sequence of SEQ ID NO: 298, a
LCDR1 comprising the sequence of SEQ ID NO: 299, a LCDR2 comprising
the sequence of SEQ ID NO: 300, a LCDR3 comprising the sequence of
SEQ ID NO: 301; cc. HCDR1 comprising the sequence of SEQ ID NO:
306, a HCDR2 comprising the sequence of SEQ ID NO: 307, a HCDR3
comprising the sequence of SEQ ID NO: 308, a LCDR1 comprising the
sequence of SEQ ID NO: 309, a LCDR2 comprising the sequence of SEQ
ID NO: 310, a LCDR3 comprising the sequence of SEQ ID NO: 311; dd.
HCDR1 comprising the sequence of SEQ ID NO: 316, a HCDR2 comprising
the sequence of SEQ ID NO: 317, a HCDR3 comprising the sequence of
SEQ ID NO: 318, a LCDR1 comprising the sequence of SEQ ID NO: 319,
a LCDR2 comprising the sequence of SEQ ID NO: 320, a LCDR3
comprising the sequence of SEQ ID NO: 321; ee. HCDR1 comprising the
sequence of SEQ ID NO: 326, a HCDR2 comprising the sequence of SEQ
ID NO: 327, a HCDR3 comprising the sequence of SEQ ID NO: 328, a
LCDR1 comprising the sequence of SEQ ID NO: 329, a LCDR2 comprising
the sequence of SEQ ID NO: 330, a LCDR3 comprising the sequence of
SEQ ID NO: 331; ff. HCDR1 comprising the sequence of SEQ ID NO:
336, a HCDR2 comprising the sequence of SEQ ID NO: 337, a HCDR3
comprising the sequence of SEQ ID NO: 338, a LCDR1 comprising the
sequence of SEQ ID NO: 339, a LCDR2 comprising the sequence of SEQ
ID NO: 340, a LCDR3 comprising the sequence of SEQ ID NO: 341; gg.
HCDR1 comprising the sequence of SEQ ID NO: 346, a HCDR2 comprising
the sequence of SEQ ID NO: 347, a HCDR3 comprising the sequence of
SEQ ID NO: 348, a LCDR1 comprising the sequence of SEQ ID NO: 349,
a LCDR2 comprising the sequence of SEQ ID NO: 350, a LCDR3
comprising the sequence of SEQ ID NO: 351; hh. HCDR1 comprising the
sequence of SEQ ID NO: 356, a HCDR2 comprising the sequence of SEQ
ID NO: 357, a HCDR3 comprising the sequence of SEQ ID NO: 358, a
LCDR1 comprising the sequence of SEQ ID NO: 359, a LCDR2 comprising
the sequence of SEQ ID NO: 360, a LCDR3 comprising the sequence of
SEQ ID NO: 361; ii. HCDR1 comprising the sequence of SEQ ID NO:
366, a HCDR2 comprising the sequence of SEQ ID NO: 367, a HCDR3
comprising the sequence of SEQ ID NO: 368, a LCDR1 comprising the
sequence of SEQ ID NO: 369, a LCDR2 comprising the sequence of SEQ
ID NO: 370, a LCDR3 comprising the sequence of SEQ ID NO: 371; jj.
HCDR1 comprising the sequence of SEQ ID NO: 376, a HCDR2 comprising
the sequence of SEQ ID NO: 377, a HCDR3 comprising the sequence of
SEQ ID NO: 378, a LCDR1 comprising the sequence of SEQ ID NO: 379,
a LCDR2 comprising the sequence of SEQ ID NO: 380, a LCDR3
comprising the sequence of SEQ ID NO: 381; kk. HCDR1 comprising the
sequence of SEQ ID NO: 386, a HCDR2 comprising the sequence of SEQ
ID NO: 387, a HCDR3 comprising the sequence of SEQ ID NO: 388, a
LCDR1 comprising the sequence of SEQ ID NO: 389, a LCDR2 comprising
the sequence of SEQ ID NO: 390, a LCDR3 comprising the sequence of
SEQ ID NO: 391; ll. HCDR1 comprising the sequence of SEQ ID NO:
396, a HCDR2 comprising the sequence of SEQ ID NO: 397, a HCDR3
comprising the sequence of SEQ ID NO: 398, a LCDR1 comprising the
sequence of SEQ ID NO: 399, a LCDR2 comprising the sequence of SEQ
ID NO: 400, a LCDR3 comprising the sequence of SEQ ID NO: 401; mm.
HCDR1 comprising the sequence of SEQ ID NO: 406, a HCDR2 comprising
the sequence of SEQ ID NO: 407, a HCDR3 comprising the sequence of
SEQ ID NO: 408, a LCDR1 comprising the sequence of SEQ ID NO: 409,
a LCDR2 comprising the sequence of SEQ ID NO: 410, a LCDR3
comprising the sequence of SEQ ID NO: 411; nn. HCDR1 comprising the
sequence of SEQ ID NO: 416, a HCDR2 comprising the sequence of SEQ
ID NO: 417, a HCDR3 comprising the sequence of SEQ ID NO: 418, a
LCDR1 comprising the sequence of SEQ ID NO: 419, a LCDR2 comprising
the sequence of SEQ ID NO: 420, a LCDR3 comprising the sequence of
SEQ ID NO: 421; oo. HCDR1 comprising the sequence of SEQ ID NO:
426, a HCDR2 comprising the sequence of SEQ ID NO: 427, a HCDR3
comprising the sequence of SEQ ID NO: 428, a LCDR1 comprising the
sequence of SEQ ID NO: 429, a LCDR2 comprising the sequence of SEQ
ID NO: 430, a LCDR3 comprising the sequence of SEQ ID NO: 431.
36. The antibody or an antigen-binding fragment thereof of claim
35, wherein said antibody or antigen-binding domain comprises a
heavy chain variable region comprises a sequence selected from the
group consisting of SEQ ID NO: 7, 17, 27, 37, 47, 57, 61, 71, 81,
91, 101, 111, 142, 152, 162, 172, 182, 192, 202, 212, 222, 232,
242, 252, 262, 272, 282, 292, 302, 312, 322, 332, 342, 352, 362,
372, 382, 392, 402, 412, 422 and 432, or a homologous sequence
thereof having at least 80% sequence identity.
37. The antibody or an antigen-binding fragment thereof of claim
35, wherein said antibody or antigen-binding domain comprises a
light chain variable region comprises a sequence selected from the
group consisting of SEQ ID NO: 8, 18, 28, 38, 48, 58, 62, 72, 82,
92, 102, 112, 143, 153, 163, 173, 183, 193, 203, 213, 223, 233,
243, 253, 263, 273, 283, 293, 303, 313, 323, 333, 343, 353, 363,
373, 383, 393, 403, 413, 423 and 433, or a homologous sequence
thereof having at least 80% sequence identity.
38. The antibody or an antigen-binding fragment thereof of claim
35, wherein said antibody or antigen-binding domain comprises a
pair of heavy chain variable region and light chain variable region
sequences selected from the group consisting of: SEQ ID NOs: 7/8,
17/18, 27/28, 37/38, 47/48, 57/58, 61/62, 71/72, 81/82, 91/92,
101/102, 111/112, and 142/143, 152/153, 162/163, 172/173, 182/183,
192/193, 202/203, 212/213, 222/223, 232/233, 242/243, 252/253,
262/263, 272/273, 282/283, 292/293, 302/303, 312/313, 322/323,
332/333, 342/343, 352/353, 362/363, 372/373, 382/383, 392/393,
402/403, 412/413, 422/423 and 432/433, or a pair of homologous
sequences thereof having at least 80% sequence identity yet
retaining specific binding affinity to RBD of spike protein of
SARS-CoV-2.
39. The antibody or an antigen-binding fragment thereof of claim
35, wherein said antibody or antigen-binding domain comprises at
least one amino acid subsequent substitutions in said
antigen-binding domain.
40. The antibody or an antigen-binding fragment thereof of claim
39, wherein said subsequent substitution comprises substituting a
cysteine residue to a non-cysteine residue.
41. The antibody or an antigen-binding fragment thereof of claim
40, wherein said cysteine residue is substituted with a serine
residue.
42. The antibody or an antigen-binding fragment thereof of claim
35, wherein said antibody or antigen-binding domain further
comprises an immunoglobulin constant region of human IgG.
43. The antibody or an antigen-binding fragment thereof of claim
42, wherein said antibody or antigen-binding domain comprises at
least one amino acid substitutions in said human IgG constant
domain, a light chain of said modified antibody, a heavy chain of
said antibody, or a combination thereof.
44. The antibody or an antigen-binding fragment thereof of claim
43, wherein said antibody or antigen-binding domain comprises at
least one amino acid substitutions in said human IgG constant
domain resulting in reduced effector functions relative to a
wildtype Fc.
45. The antibody or an antigen-binding fragment thereof of claim
44, wherein said antibody or antigen-binding domain comprises one
or more amino acid substitution(s) at a position selected from the
group consisting of: 220, 226, 229, 233, 234, 235, 236, 237, 238,
267, 268, 269, 270, 297, 309, 318, 320, 322, 325, 328, 329, 330,
and 331 of the Fc region, wherein the numbering of the residues in
the Fc region is that of the EU index as in Kabat.
46. The antibody or an antigen-binding fragment thereof of claim
45, wherein said antibody or antigen-binding domain comprises one
or more amino acid substitution(s) selected from the group
consisting of 220S, 226S, 228P, 229S, 233P, 234V, 234G, 234A, 234F,
234A, 235A, 235G, 235E, 236E, 236R, 237A, 237K, 238S, 267R, 268A,
268Q, 269R, 297A, 297Q, 297G, 309L, 318A, 322A, 325L, 328R, 330S,
331S and any combination thereof.
47. The antibody or an antigen-binding fragment thereof of claim
45, wherein said antibody or antigen-binding domain comprises a
combination of mutations selected from the group consisting of: a)
K322A, L234A, and L235A; b) P331S, L234F, and L235E; c) L234A and
L235A; c) N297A; d) N297Q; e) N297G; f) L235E; g) L234A and L235A
(IgG1); h) F234A and L235A (IgG4); i) H268Q, V309L, A330S and P331S
(IgG2); j) V234A, G237A, P238S, H268A, V309L, A330S and P331S
(IgG2).
48. The antibody or an antigen-binding fragment thereof of claim
43, wherein said antibody or antigen-binding domain comprises one
or more amino acid residue modifications or substitutions resulting
in improved binding affinity to neonatal Fc receptor (FcRn) at pH
6.0 while retaining minimal binding at pH 7.4, or increased serum
half life of the antibody.
49. The antibody or an antigen-binding fragment thereof of claim
48, wherein said antibody or antigen-binding domain comprises one
or more amino acid substitution(s) at a position selected from the
group consisting of: 234, 235, 238, 250, 252, 254, 256; 259; 272,
305, 307, 308, 311, 312, 322, 328, 331, 378, 380, 382, 428, 432,
433, 434, 435, 436 and 437, all positions by EU numbering.
50. The antibody or an antigen-binding fragment thereof of claim
49, wherein said antibody or antigen-binding domain comprises one
or more amino acid substitution(s) selected from the group
consisting of 234F, 235Q, 238D, 250Q, 252T, 252Y, 254T, 256E, 259I,
272A, 305A, 307A, 308F, 311A, 322Q, 328E, 331S, 380A, 428L, 432C,
433K, 433S, 434S, 434Y, 434F, 434W, 434A, 435H, 436L, 437C and any
combination thereof.
51. The antibody or an antigen-binding fragment thereof of claim
50, wherein said antibody or antigen-binding domain has increased
serum half-life or improved pH-dependent binding to FcRn and
comprises a combination of mutations selected from the group
consisting of: a) M428L and N434S; b) P238D and L328E; c) M252Y,
S254T and T256E; d) L234F, L235Q, K322Q, M252T, S254T and T256E; e)
M428L, V259I and V308F; f) H433K and N434Y; g) H433K and N434F; h)
T250Q and M428L; i) T307A, E380A and N434A; and j) 432C, 433S,
434W, 435H, 436L, and 437C.
52. The antibody or an antigen-binding fragment thereof of claim
34, wherein said modified antibody or said antigen-binding fragment
thereof is a diabody, a Fab, a Fab', a F(ab')2, a Fd, an Fv
fragment, a disulfide stabilized Fv fragment, a (dsFv)2, a
bispecific dsFv, a disulfide stabilized diabody, a single-chain
antibody molecule, an scFv dimer, a bispecific scFv dimer, or a
multispecific antibody.
53. The antibody or an antigen-binding fragment thereof of claim
35, which is linked to one or more conjugate moieties.
54. A bispecific antibody comprising one or more of the
antigen-binding fragment of claim 35.
55. An isolated polynucleotide encoding the antibody or antigen
binding fragment of claim 35.
56. A vector comprising the isolated polynucleotide of claim
55.
57. A host cell comprising the vector of claim 56.
58. A pharmaceutical composition comprising at least one said
antibody or an antigen-binding fragment thereof of claim 35, at
least one nucleic acid encoding said modified antibody or said
antigen-binding fragment thereof, or a combination thereof, and one
or more pharmaceutically acceptable carriers.
59. A method for treating or preventing a disease in a subject in
need thereof, said method comprising administering an effective
dosage of said pharmaceutical composition of claim 58 to said
subject.
60. The method of claim 59, wherein the disease is associated with
SARs-CoV-2 infection.
61. The method of claim 59, wherein said pharmaceutical composition
is administered to said subject having no symptoms or free from
known infections of said SARS-CoV-2, prior to said subject being
infected with said SARS-CoV-2, prior to said subject exhibiting any
symptoms of the infection of said SARS-CoV-2, or a combination
thereof.
62. The method of claims 59, wherein said pharmaceutical
composition is administered to said subject via intravenous
injection (IV), intramuscular injection (IM), subcutaneous (SC)
injection, or a combination thereof.
63. The method of claim 59, wherein said subject is a patient of
age 60, 70 or 80 years old or older.
64. The method of claim 59, wherein said effective dosage is
determined by a dosing process that comprises determining
concentration progression data based on calculated or measured
pharmacokinetics (PK), testing plasma concentrations over a testing
period of time, predicted plasma concentrations over a prediction
period of time, or a combination thereof, of said antibody or said
antigen-binding fragment thereof, and producing said effective
dosage based on said concentration progression data.
65. The method of claim 59, further comprising administering a
pharmaceutically effective amount of one or more bioactive agents
to said subject simultaneously or sequentially with said
pharmaceutical composition, wherein said bioactive agent comprises
a therapeutic agent or a prophylactic agent selected from an
anti-viral agent, an antiviral peptide, an anti-viral antibody, an
anti-viral compound, an anti-viral cytokine, an anti-viral
oligonucleotide, an RNA dependent RNA polymerase inhibitor, a
non-nucleoside reverse transcriptase inhibitor (NNRTI), nucleoside
reverse transcriptase inhibitor (NRTI), purine nucleoside,
antiviral interferon, adamantine antiviral compound, remdesivir,
chloroquine, hydroxychloroquine, lopinavir, ritonavir, APN01,
favilavir, mesalazine, toremifene, eplerenone, paroxetine,
sirolimus, dactinomycin, irbesartan, emodin, mercaptopurine,
melatonin, quinacrine, carvedilol, colchicine, camphor, equilin,
oxymetholone, nafamosta, camostat, baricitinib, darunavir,
ribavirin, galidesivir, BCX-4430, Arbidol, nitazoxanide, one or
more derivatives thereof, or any combination thereof.
Description
CROSS REFERENCE
[0001] This application is a continuation application of U.S.
application Ser. No. 16/953,304, and claims the priorities of
Foreign Applications No. CN202010203065.1, filed on Mar. 20, 2020;
PCT/CN2020/080532, filed Mar. 21, 2020; PCT/CN2020/084097, filed on
Apr. 10, 2020; PCT/CN2020/084805, filed on Apr. 14, 2020; and
PCT/CN2020/108718, filed on Aug. 12, 2020; which are hereby
incorporated by reference.
FIELD OF THE DISCLOSURE
[0002] The present disclosure generally relates to novel
anti-SARS-COV-2 antibodies, pharmaceutical composition containing
the same and the use thereof.
BACKGROUND
[0003] The recent outbreak of the new coronavirus, SARS-CoV-2 poses
a serious global health emergency. SARS-CoV-2 is a positive-sense
single-stranded RNA (+ssRNA) virus which belongs to the
betacoronavirus family and shares substantial genetic and
functional similarity with other pathogenic human
betacoronaviruses, including Severe Acute Respiratory Syndrome
Coronavirus (SARS-CoV, also called SARS-CoV-1) and Middle East
Respiratory Syndrome Coronavirus (MERS-CoV). Like other
coronaviruses, SARS-CoV-2 has four structural proteins, known as
the S (spike), E (envelope), M (membrane), and N (nucleocapsid)
proteins; the S, E, and M proteins together create the viral
envelope; inside the envelope is the N protein bounding to the RNA
genome (.about.30 kb) in a continuous beads-on-a-string type
conformation.
[0004] The spike protein is the protein responsible for allowing
the SARS-CoV-2 virus to attach to the membrane of a host cell, the
receptor binding domain (RBD) of the spike protein of SARS-CoV-2
recognizes and attaches to the angiotensin-converting enzyme 2
(ACE2) receptor of host cells to use them as a mechanism of cell
entry. The overall ACE2-binding mechanism is virtually the same
between SARS-CoV-2 RBD and SARS-CoV RBD, indicating convergent
ACE2-binding evolution between these two viruses. This suggests
that disruption of the RBD and ACE2 interaction would block the
entry of SARS-CoV-2 into the target cell. Indeed, a few such
disruptive agents targeted to ACE2 have been shown to inhibit
SARS-CoV infection. However, given the important physiological
roles of ACE2 in vivo, these agents may have undesired side
effects. Anti-RBD antibodies, on the other hand, are therefore more
favorable. Furthermore, SARS-CoV-RBD or MERS-CoV RBD-based vaccine
studies in experimental animals have also shown strong polyclonal
antibody responses that inhibit viral entry. Such critical
proof-of-concept findings indicate that anti-RBD antibodies might
effectively block SARS-CoV-2 entry.
[0005] No SARS-CoV-2-specific treatments or vaccine are currently
available, and the currently existing detective measures for
SARS-CoV-2 infection are time-consuming and insensitive. Hence,
there is an urgent need for novel anti-SARS-CoV-2 antibodies.
BRIEF SUMMARY OF THE INVENTION
[0006] In one aspect, the present disclosure is directed to a
modified antibody or an antigen-binding fragment thereof comprising
at least an antigen-binding domain having an antigen-binding
affinity and a covalently linked modified human IgG constant
domain, wherein the antigen-binding affinity comprises SARS-CoV-2
binding affinity, the antigen-binding affinity comprises at least
50% less or non-detectable binding affinity to SARS-CoV or MERS-CoV
compared to the SARS-CoV-2 binding affinity, and wherein the
modified human IgG constant domain comprises a substitution with
tyrosine at amino acid residue 252, a substitution with threonine
at amino acid residue 254, and a substitution with glutamic acid at
amino acid residue 256, numbered according to the EU index as in
Kabat, the modified antibody has an increased affinity for FcRn
compared to the affinity to FcRn of an antibody having a wild type
human IgG constant domain.
[0007] In another aspect, the present disclosure is directed to a
pharmaceutical composition comprising at least one the modified
antibody or an antigen-binding fragment thereof of disclosed
herein, at least one nucleic acid encoding the modified antibody or
the antigen-binding fragment thereof, or a combination thereof, and
one or more pharmaceutically acceptable carriers.
[0008] In another aspect, the present disclosure is directed to a
method for treating or preventing a disease in a subject in need
thereof, the method comprising administering an effective dosage of
any of the pharmaceutical composition of disclosed herein to the
subject;
[0009] wherein the pharmaceutical composition is configured to be
administered to the subject to maintain a plasma concentration of
the modified antibody or an antigen-binding fragment thereof in a
therapeutic effective range of from 10 .mu.g/mL to 3500 .mu.g/mL
for a time period in a range of from 1 day to 12 months after
administering the pharmaceutical composition; and
[0010] wherein the subject is infected with, exhibiting one or more
symptoms of being infected with, or at risk of being infected with
the SARS-CoV-2.
[0011] In another aspect, the present disclosure provides an
isolated or recombinant antibody or an antigen-binding fragment
thereof, which is capable of specifically binding to SARS-CoV-2,
and exhibiting at least 50% less binding or non-detectable binding
to SARS-CoV or MERS-CoV.
[0012] In another aspect, the present disclosure provides an
isolated or recombinant antibody or an antigen-binding fragment
thereof, having one or more features selected from the group
consisting of: a) capable of specifically binding to spike protein
of SARS-CoV-2 and exhibiting at least 50% less binding to spike
protein of SARS-CoV or spike protein of MERS-CoV; b) capable of
specifically binding to receptor-binding domain (RBD) of the spike
protein of SARS-CoV-2 comprising the amino acid sequence of SEQ ID
NO: 128; c) exhibiting binding to RBD of spike protein of SARS-CoV
comprising the amino acid sequence of SEQ ID NO: 124 at a level
that is non-detectable or that is no more than 50% of the binding
to the RBD of spike protein of SARS-CoV-2; d) exhibiting binding to
RBD of spike protein of MERS-CoV comprising the amino acid sequence
of SEQ ID NO: 126 at a level that is non-detectable or that is no
more than 50% of the binding to RBD of the spike protein of
SARS-CoV-2; e) capable of binding to the RBD of spike protein of
SARS-CoV-2 at a K.sub.d value of no more than 1.times.10.sup.-7M as
measured by Surface Plasmon resonance (SPR); f) exhibiting binding
to the RBD of spike protein of SARS-CoV or the RBD of spike protein
of MERS-CoV at a K.sub.d value of at least 1.times.10.sup.-6 M as
measured by SPR; g) capable of exhibiting at least 30% competition
at with 2 .mu.M angiotensin converting enzyme 2 (ACE2) receptor,
for binding to the RBD of spike protein of SARS-CoV-2 immobilized
at a resonance units (RU) of 250, as measured by SPR; h) capable of
binding to the RBD of spike protein of SARS-CoV-2 at an
neutralizing activity at an IC.sub.50 value of no more than 100
.mu.g/ml (for example, no more than 50 .mu.g/ml, no more than 40
.mu.g/ml, no more than 30 .mu.g/ml, no more than 25 .mu.g/ml, no
more than 20 .mu.g/ml, no more than 15 .mu.g/ml, no more than 10
.mu.g/ml, no more than 8 .mu.g/ml, no more than 6 .mu.g/ml, no more
than 4 .mu.g/ml, no more than 2 .mu.g/ml, or no more than 1
.mu.g/ml), as measured by pseudovirus neutralization assay, and 1)
capable of binding to the RBD of spike protein of SARS-CoV-2 at an
neutralizing activity at an IC.sub.50 value of no more than 1
.mu.g/ml (for example, no more than 50 ng/ml, no more than 40
ng/ml, no more than 30 ng/ml, no more than 25 ng/ml, no more than
20 ng/ml, no more than 15 ng/ml, no more than 10 ng/ml, no more
than 8 ng/ml, no more than 6 ng/ml, no more than 4 ng/ml, no more
than 2 ng/ml, or no more than 1 ng/ml), as measured by live virus
neutralization assay using focus reduction neutralization test
(FRNT) method.
[0013] In yet another aspect, the present disclosure provides an
isolated or recombinant antibody or an antigen-binding fragment
thereof capable of specifically binding to RBD of spike protein of
SARS-CoV-2.
[0014] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID
NO: 3.
[0015] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 11, SEQ ID NO: 12, and SEQ
ID NO: 13.
[0016] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 21, SEQ ID NO: 22, and SEQ
ID NO: 23.
[0017] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 31, SEQ ID NO: 32, and SEQ
ID NO: 33.
[0018] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 41, SEQ ID NO: 42, and SEQ
ID NO: 43.
[0019] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 51, SEQ ID NO: 52, and SEQ
ID NO: 53.
[0020] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 65, SEQ ID NO: 66, and SEQ
ID NO: 67.
[0021] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 75, SEQ ID NO: 76, and SEQ
ID NO: 77.
[0022] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 85, SEQ ID NO: 86, and SEQ
ID NO: 87.
[0023] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 95, SEQ ID NO: 96, and SEQ
ID NO: 97.
[0024] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 105, SEQ ID NO: 106, and SEQ
ID NO: 107.
[0025] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 136, SEQ ID NO: 137, and SEQ
ID NO: 138.
[0026] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 146, SEQ ID NO: 147, and SEQ
ID NO: 148.
[0027] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 156, SEQ ID NO: 157, and SEQ
ID NO: 158.
[0028] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 166, SEQ ID NO: 167, and SEQ
ID NO: 168.
[0029] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 176, SEQ ID NO: 177, and SEQ
ID NO: 178.
[0030] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 186, SEQ ID NO: 187, and SEQ
ID NO: 188.
[0031] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 196, SEQ ID NO: 197, and SEQ
ID NO: 198.
[0032] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 206, SEQ ID NO: 207, and SEQ
ID NO: 208.
[0033] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 216, SEQ ID NO: 217, and SEQ
ID NO: 218.
[0034] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 226, SEQ ID NO: 227, and SEQ
ID NO: 228.
[0035] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 236, SEQ ID NO: 237, and SEQ
ID NO: 238.
[0036] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 246, SEQ ID NO: 247, and SEQ
ID NO: 248.
[0037] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 256, SEQ ID NO: 257, and SEQ
ID NO: 258.
[0038] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 266, SEQ ID NO: 267, and SEQ
ID NO: 268.
[0039] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 276, SEQ ID NO: 277, and SEQ
ID NO: 278.
[0040] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 286, SEQ ID NO: 287, and SEQ
ID NO: 288.
[0041] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 296, SEQ ID NO: 297, and SEQ
ID NO: 298.
[0042] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 306, SEQ ID NO: 307, and SEQ
ID NO: 308.
[0043] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 316, SEQ ID NO: 317, and SEQ
ID NO: 318.
[0044] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 326, SEQ ID NO: 327, and SEQ
ID NO: 328.
[0045] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 336, SEQ ID NO: 337, and SEQ
ID NO: 338.
[0046] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 346, SEQ ID NO: 347, and SEQ
ID NO: 348.
[0047] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 266, SEQ ID NO: 267, and SEQ
ID NO: 268.
[0048] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 356, SEQ ID NO: 357, and SEQ
ID NO: 358.
[0049] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 366, SEQ ID NO: 367, and SEQ
ID NO: 368.
[0050] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 376, SEQ ID NO: 377, and SEQ
ID NO: 378.
[0051] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 386, SEQ ID NO: 387, and SEQ
ID NO: 388.
[0052] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 396, SEQ ID NO: 397, and SEQ
ID NO: 398.
[0053] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 406, SEQ ID NO: 407, and SEQ
ID NO: 408.
[0054] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 416, SEQ ID NO: 417, and SEQ
ID NO: 418.
[0055] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 heavy chain
CDR sequences selected from SEQ ID NO: 426, SEQ ID NO: 427, and SEQ
ID NO: 428.
[0056] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID
NO: 6.
[0057] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 14, SEQ ID NO: 15, and SEQ
ID NO: 16.
[0058] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 24, SEQ ID NO: 25 and SEQ ID
NO: 26.
[0059] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID
NO: 36.
[0060] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 44, SEQ ID NO: 45 and SEQ ID
NO: 46.
[0061] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID
NO: 56.
[0062] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 68, SEQ ID NO: 69, and SEQ
ID NO: 70.
[0063] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 78, SEQ ID NO: 79, and SEQ
ID NO: 80.
[0064] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 88, SEQ ID NO: 89, and SEQ
ID NO: 90.
[0065] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 98, SEQ ID NO: 99, and SEQ
ID NO: 100.
[0066] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 108, SEQ ID NO: 109, and SEQ
ID NO: 110.
[0067] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 139, SEQ ID NO: 140, and SEQ
ID NO: 141.
[0068] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 149, SEQ ID NO: 150, and SEQ
ID NO: 151.
[0069] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 159, SEQ ID NO: 160, and SEQ
ID NO: 161.
[0070] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 169, SEQ ID NO: 170, and SEQ
ID NO: 171.
[0071] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 179, SEQ ID NO: 180, and SEQ
ID NO: 181.
[0072] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 189, SEQ ID NO: 190, and SEQ
ID NO: 191.
[0073] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 199, SEQ ID NO: 200, and SEQ
ID NO: 201.
[0074] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 209, SEQ ID NO: 210, and SEQ
ID NO: 211.
[0075] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 219, SEQ ID NO: 220, and SEQ
ID NO: 221.
[0076] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 229, SEQ ID NO: 230, and SEQ
ID NO: 231.
[0077] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 239, SEQ ID NO: 240, and SEQ
ID NO: 241.
[0078] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 249, SEQ ID NO: 250, and SEQ
ID NO: 251.
[0079] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 259, SEQ ID NO: 260, and SEQ
ID NO: 261.
[0080] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 269, SEQ ID NO: 270, and SEQ
ID NO: 271.
[0081] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 279, SEQ ID NO: 280, and SEQ
ID NO: 281.
[0082] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 289, SEQ ID NO: 290, and SEQ
ID NO: 291.
[0083] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 299, SEQ ID NO: 300, and SEQ
ID NO: 301.
[0084] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 309, SEQ ID NO: 310, and SEQ
ID NO: 311.
[0085] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 319, SEQ ID NO: 320, and SEQ
ID NO: 321.
[0086] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 329, SEQ ID NO: 330, and SEQ
ID NO: 331.
[0087] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 339, SEQ ID NO: 340, and SEQ
ID NO: 341.
[0088] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 349, SEQ ID NO: 350, and SEQ
ID NO: 351.
[0089] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 359, SEQ ID NO: 360, and SEQ
ID NO: 361.
[0090] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 369, SEQ ID NO: 370, and SEQ
ID NO: 371.
[0091] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 379, SEQ ID NO: 380, and SEQ
ID NO: 381.
[0092] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 389, SEQ ID NO: 390, and SEQ
ID NO: 391.
[0093] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 399, SEQ ID NO: 400, and SEQ
ID NO: 401.
[0094] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 409, SEQ ID NO: 410, and SEQ
ID NO: 411.
[0095] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 419, SEQ ID NO: 420, and SEQ
ID NO: 421.
[0096] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 429, SEQ ID NO: 430, and SEQ
ID NO: 431.
[0097] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a heavy chain CDR1
(HCDR1) comprising the sequence of SEQ ID NO: 1, a heavy chain CDR2
(HCDR2) comprising the sequence of SEQ ID NO: 2, a heavy chain CDR3
(HCDR3) comprising the sequence of SEQ ID NO: 3; a light chain CDR1
(LCDR1) comprising the sequence of SEQ ID NO: 4, a light chain CDR2
(LCDR2) comprising the sequence of SEQ ID NO: 5, and a light chain
CDR3 (LCDR3) comprising the sequence of SEQ ID NO: 6.
[0098] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 11, a HCDR2 comprising the sequence of SEQ
ID NO: 12, a HCDR3 comprising the sequence of SEQ ID NO: 13, a
LCDR1 comprising the sequence of SEQ ID NO: 14, a LCDR2 comprising
the sequence of SEQ ID NO: 15, and a LCDR3 comprising the sequence
of SEQ ID NO: 16.
[0099] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 21, a HCDR2 comprising the sequence of SEQ
ID NO: 22, a HCDR3 comprising the sequence of SEQ ID NO: 23, a
LCDR1 comprising the sequence of SEQ ID NO: 24, a LCDR2 comprising
the sequence of SEQ ID NO: 25, and a LCDR3 comprising the sequence
of SEQ ID NO: 26.
[0100] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 31, a HCDR2 comprising the sequence of SEQ
ID NO: 32, a HCDR3 comprising the sequence of SEQ ID NO: 33, a
LCDR1 comprising the sequence of SEQ ID NO: 34, a LCDR2 comprising
the sequence of SEQ ID NO: 35, and a LCDR3 comprising the sequence
of SEQ ID NO: 36.
[0101] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 41, a HCDR2 comprising the sequence of SEQ
ID NO: 42, a HCDR3 comprising the sequence of SEQ ID NO: 43, a
LCDR1 comprising the sequence of SEQ ID NO: 44, a LCDR2 comprising
the sequence of SEQ ID NO: 45, and a LCDR3 comprising the sequence
of SEQ ID NO: 46.
[0102] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 51, a HCDR2 comprising the sequence of SEQ
ID NO: 52, a HCDR3 comprising the sequence of SEQ ID NO: 53, a
LCDR1 comprising the sequence of SEQ ID NO: 54, a LCDR2 comprising
the sequence of SEQ ID NO: 55, and a LCDR3 comprising the sequence
of SEQ ID NO: 56.
[0103] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 65, a HCDR2 comprising the sequence of SEQ
ID NO: 66, a HCDR3 comprising the sequence of SEQ ID NO: 67, a
LCDR1 comprising the sequence of SEQ ID NO: 68, a LCDR2 comprising
the sequence of SEQ ID NO: 69, and a LCDR3 comprising the sequence
of SEQ ID NO: 70.
[0104] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 75, a HCDR2 comprising the sequence of SEQ
ID NO: 76, a HCDR3 comprising the sequence of SEQ ID NO: 77, a
LCDR1 comprising the sequence of SEQ ID NO: 78, a LCDR2 comprising
the sequence of SEQ ID NO: 79, and a LCDR3 comprising the sequence
of SEQ ID NO: 80.
[0105] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 85, a HCDR2 comprising the sequence of SEQ
ID NO: 86, a HCDR3 comprising the sequence of SEQ ID NO: 87, a
LCDR1 comprising the sequence of SEQ ID NO: 88, a LCDR2 comprising
the sequence of SEQ ID NO: 89, and a LCDR3 comprising the sequence
of SEQ ID NO: 90.
[0106] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 95, a HCDR2 comprising the sequence of SEQ
ID NO: 96, a HCDR3 comprising the sequence of SEQ ID NO: 97, a
LCDR1 comprising the sequence of SEQ ID NO: 98, a LCDR2 comprising
the sequence of SEQ ID NO: 99, and a LCDR3 comprising the sequence
of SEQ ID NO: 100.
[0107] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 105, a HCDR2 comprising the sequence of SEQ
ID NO: 106, a HCDR3 comprising the sequence of SEQ ID NO: 107, a
LCDR1 comprising the sequence of SEQ ID NO: 108, a LCDR2 comprising
the sequence of SEQ ID NO: 109, and a LCDR3 comprising the sequence
of SEQ ID NO: 110.
[0108] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 136, a HCDR2 comprising the sequence of SEQ
ID NO: 137, a HCDR3 comprising the sequence of SEQ ID NO: 138, a
LCDR1 comprising the sequence of SEQ ID NO: 139, a LCDR2 comprising
the sequence of SEQ ID NO: 140, and a LCDR3 comprising the sequence
of SEQ ID NO: 141.
[0109] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 146, a HCDR2 comprising the sequence of SEQ
ID NO: 147, a HCDR3 comprising the sequence of SEQ ID NO: 148, a
LCDR1 comprising the sequence of SEQ ID NO: 149, a LCDR2 comprising
the sequence of SEQ ID NO: 150, and a LCDR3 comprising the sequence
of SEQ ID NO: 151.
[0110] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 156, a HCDR2 comprising the sequence of SEQ
ID NO: 157, a HCDR3 comprising the sequence of SEQ ID NO: 158, a
LCDR1 comprising the sequence of SEQ ID NO: 159, a LCDR2 comprising
the sequence of SEQ ID NO: 160, and a LCDR3 comprising the sequence
of SEQ ID NO: 161.
[0111] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 166, a HCDR2 comprising the sequence of SEQ
ID NO: 167, a HCDR3 comprising the sequence of SEQ ID NO: 168, a
LCDR1 comprising the sequence of SEQ ID NO: 169, a LCDR2 comprising
the sequence of SEQ ID NO: 170, and a LCDR3 comprising the sequence
of SEQ ID NO: 171.
[0112] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 176, a HCDR2 comprising the sequence of SEQ
ID NO: 177, a HCDR3 comprising the sequence of SEQ ID NO: 178, a
LCDR1 comprising the sequence of SEQ ID NO: 179, a LCDR2 comprising
the sequence of SEQ ID NO: 180, and a LCDR3 comprising the sequence
of SEQ ID NO: 181.
[0113] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 186, a HCDR2 comprising the sequence of SEQ
ID NO: 187, a HCDR3 comprising the sequence of SEQ ID NO: 188, a
LCDR1 comprising the sequence of SEQ ID NO: 189, a LCDR2 comprising
the sequence of SEQ ID NO: 190, and a LCDR3 comprising the sequence
of SEQ ID NO: 191.
[0114] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 196, a HCDR2 comprising the sequence of SEQ
ID NO: 197, a HCDR3 comprising the sequence of SEQ ID NO: 198, a
LCDR1 comprising the sequence of SEQ ID NO: 199, a LCDR2 comprising
the sequence of SEQ ID NO: 200, and a LCDR3 comprising the sequence
of SEQ ID NO: 201.
[0115] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 206, a HCDR2 comprising the sequence of SEQ
ID NO: 207, a HCDR3 comprising the sequence of SEQ ID NO: 208, a
LCDR1 comprising the sequence of SEQ ID NO: 209, a LCDR2 comprising
the sequence of SEQ ID NO: 210, and a LCDR3 comprising the sequence
of SEQ ID NO: 211.
[0116] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 216, a HCDR2 comprising the sequence of SEQ
ID NO: 217, a HCDR3 comprising the sequence of SEQ ID NO: 218, a
LCDR1 comprising the sequence of SEQ ID NO: 219, a LCDR2 comprising
the sequence of SEQ ID NO: 220, and a LCDR3 comprising the sequence
of SEQ ID NO: 221.
[0117] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 226, a HCDR2 comprising the sequence of SEQ
ID NO: 227, a HCDR3 comprising the sequence of SEQ ID NO: 228, a
LCDR1 comprising the sequence of SEQ ID NO: 229, a LCDR2 comprising
the sequence of SEQ ID NO: 230, and a LCDR3 comprising the sequence
of SEQ ID NO: 231.
[0118] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 236, a HCDR2 comprising the sequence of SEQ
ID NO: 237, a HCDR3 comprising the sequence of SEQ ID NO: 238, a
LCDR1 comprising the sequence of SEQ ID NO: 239, a LCDR2 comprising
the sequence of SEQ ID NO: 240, and a LCDR3 comprising the sequence
of SEQ ID NO: 241.
[0119] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 246, a HCDR2 comprising the sequence of SEQ
ID NO: 247, a HCDR3 comprising the sequence of SEQ ID NO: 248, a
LCDR1 comprising the sequence of SEQ ID NO: 249, a LCDR2 comprising
the sequence of SEQ ID NO: 250, and a LCDR3 comprising the sequence
of SEQ ID NO: 251.
[0120] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 256, a HCDR2 comprising the sequence of SEQ
ID NO: 257, a HCDR3 comprising the sequence of SEQ ID NO: 258, a
LCDR1 comprising the sequence of SEQ ID NO: 259, a LCDR2 comprising
the sequence of SEQ ID NO: 260, and a LCDR3 comprising the sequence
of SEQ ID NO: 261.
[0121] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 266, a HCDR2 comprising the sequence of SEQ
ID NO: 267, a HCDR3 comprising the sequence of SEQ ID NO: 268, a
LCDR1 comprising the sequence of SEQ ID NO: 269, a LCDR2 comprising
the sequence of SEQ ID NO: 270, and a LCDR3 comprising the sequence
of SEQ ID NO: 271.
[0122] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 276, a HCDR2 comprising the sequence of SEQ
ID NO: 277, a HCDR3 comprising the sequence of SEQ ID NO: 278, a
LCDR1 comprising the sequence of SEQ ID NO: 279, a LCDR2 comprising
the sequence of SEQ ID NO: 280, and a LCDR3 comprising the sequence
of SEQ ID NO: 281.
[0123] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 286, a HCDR2 comprising the sequence of SEQ
ID NO: 287, a HCDR3 comprising the sequence of SEQ ID NO: 288, a
LCDR1 comprising the sequence of SEQ ID NO: 289, a LCDR2 comprising
the sequence of SEQ ID NO: 290, and a LCDR3 comprising the sequence
of SEQ ID NO: 291.
[0124] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 296, a HCDR2 comprising the sequence of SEQ
ID NO: 297, a HCDR3 comprising the sequence of SEQ ID NO: 298, a
LCDR1 comprising the sequence of SEQ ID NO: 299, a LCDR2 comprising
the sequence of SEQ ID NO: 300, and a LCDR3 comprising the sequence
of SEQ ID NO: 301.
[0125] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 306, a HCDR2 comprising the sequence of SEQ
ID NO: 307, a HCDR3 comprising the sequence of SEQ ID NO: 308, a
LCDR1 comprising the sequence of SEQ ID NO: 309, a LCDR2 comprising
the sequence of SEQ ID NO: 310, and a LCDR3 comprising the sequence
of SEQ ID NO: 311.
[0126] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 316, a HCDR2 comprising the sequence of SEQ
ID NO: 317, a HCDR3 comprising the sequence of SEQ ID NO: 318, a
LCDR1 comprising the sequence of SEQ ID NO: 319, a LCDR2 comprising
the sequence of SEQ ID NO: 320, and a LCDR3 comprising the sequence
of SEQ ID NO: 321.
[0127] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 326, a HCDR2 comprising the sequence of SEQ
ID NO: 327, a HCDR3 comprising the sequence of SEQ ID NO: 328, a
LCDR1 comprising the sequence of SEQ ID NO: 329, a LCDR2 comprising
the sequence of SEQ ID NO: 330, and a LCDR3 comprising the sequence
of SEQ ID NO: 331.
[0128] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 336, a HCDR2 comprising the sequence of SEQ
ID NO: 337, a HCDR3 comprising the sequence of SEQ ID NO: 338, a
LCDR1 comprising the sequence of SEQ ID NO: 339, a LCDR2 comprising
the sequence of SEQ ID NO: 340, and a LCDR3 comprising the sequence
of SEQ ID NO: 341.
[0129] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 346, a HCDR2 comprising the sequence of SEQ
ID NO: 347, a HCDR3 comprising the sequence of SEQ ID NO: 348, a
LCDR1 comprising the sequence of SEQ ID NO: 349, a LCDR2 comprising
the sequence of SEQ ID NO: 350, and a LCDR3 comprising the sequence
of SEQ ID NO: 351.
[0130] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 356, a HCDR2 comprising the sequence of SEQ
ID NO: 357, a HCDR3 comprising the sequence of SEQ ID NO: 358, a
LCDR1 comprising the sequence of SEQ ID NO: 359, a LCDR2 comprising
the sequence of SEQ ID NO: 360, and a LCDR3 comprising the sequence
of SEQ ID NO: 361.
[0131] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 366, a HCDR2 comprising the sequence of SEQ
ID NO: 367, a HCDR3 comprising the sequence of SEQ ID NO: 368, a
LCDR1 comprising the sequence of SEQ ID NO: 369, a LCDR2 comprising
the sequence of SEQ ID NO: 370, and a LCDR3 comprising the sequence
of SEQ ID NO: 371.
[0132] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 376, a HCDR2 comprising the sequence of SEQ
ID NO: 377, a HCDR3 comprising the sequence of SEQ ID NO: 378, a
LCDR1 comprising the sequence of SEQ ID NO: 379, a LCDR2 comprising
the sequence of SEQ ID NO: 380, and a LCDR3 comprising the sequence
of SEQ ID NO: 381.
[0133] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 386, a HCDR2 comprising the sequence of SEQ
ID NO: 387, a HCDR3 comprising the sequence of SEQ ID NO: 388, a
LCDR1 comprising the sequence of SEQ ID NO: 389, a LCDR2 comprising
the sequence of SEQ ID NO: 390, and a LCDR3 comprising the sequence
of SEQ ID NO: 391.
[0134] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 396, a HCDR2 comprising the sequence of SEQ
ID NO: 397, a HCDR3 comprising the sequence of SEQ ID NO: 398, a
LCDR1 comprising the sequence of SEQ ID NO: 399, a LCDR2 comprising
the sequence of SEQ ID NO: 400, and a LCDR3 comprising the sequence
of SEQ ID NO: 401.
[0135] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 406, a HCDR2 comprising the sequence of SEQ
ID NO: 407, a HCDR3 comprising the sequence of SEQ ID NO: 408, a
LCDR1 comprising the sequence of SEQ ID NO: 409, a LCDR2 comprising
the sequence of SEQ ID NO: 410, and a LCDR3 comprising the sequence
of SEQ ID NO: 411.
[0136] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 416, a HCDR2 comprising the sequence of SEQ
ID NO: 417, a HCDR3 comprising the sequence of SEQ ID NO: 418, a
LCDR1 comprising the sequence of SEQ ID NO: 419, a LCDR2 comprising
the sequence of SEQ ID NO: 420, and a LCDR3 comprising the sequence
of SEQ ID NO: 421.
[0137] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a HCDR1 comprising the
sequence of SEQ ID NO: 426, a HCDR2 comprising the sequence of SEQ
ID NO: 427, a HCDR3 comprising the sequence of SEQ ID NO: 428, a
LCDR1 comprising the sequence of SEQ ID NO: 429, a LCDR2 comprising
the sequence of SEQ ID NO: 430, and a LCDR3 comprising the sequence
of SEQ ID NO: 431.
[0138] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a heavy chain variable
region comprises a sequence selected from the group consisting of
SEQ ID NO: 7, 17, 27, 37, 47, 57, 61, 71, 81, 91, 101, 111, 142,
152, 162, 172, 182, 192, 202, 212, 222, 232, 242, 252, 262, 272,
282, 292, 302, 312, 322, 332, 342, 352, 362, 372, 382, 392, 402,
412, 422 and 432, or a homologous sequence thereof having at least
80% sequence identity.
[0139] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a light chain variable
region comprises a sequence selected from the group consisting of
SEQ ID NO: 8, 18, 28, 38, 48, 58, 62, 72, 82, 92, 102, 112, 143,
153, 163, 173, 183, 193, 203, 213, 223, 233, 243, 253, 263, 273,
283, 293, 303, 313, 323, 333, 343, 353, 363, 373, 383, 393, 403,
413, 423 and 433, or a homologous sequence thereof having at least
80% sequence identity.
[0140] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a pair of heavy chain
variable region and light chain variable region sequences selected
from the group consisting of: SEQ ID NOs: 7/8, 17/18, 27/28, 37/38,
47/48, 57/58, 61/62, 71/72, 81/82, 91/92, 101/102, 111/112, and
142/143, 152/153, 162/163, 172/173, 182/183, 192/193, 202/203,
212/213, 222/223, 232/233, 242/243, 252/253, 262/263, 272/273,
282/283, 292/293, 302/303, 312/313, 322/323, 332/333, 342/343,
352/353, 362/363, 372/373, 382/383, 392/393, 402/403, 412/413,
422/423 and 432/433, or a pair of homologous sequences thereof
having at least 80% sequence identity yet retaining specific
binding affinity to RBD of spike protein of SARS-CoV-2.
[0141] In some embodiments, the antibody or antigen binding
fragment of the present disclosure further comprises an
immunoglobulin constant region. In some embodiments, the
immunoglobulin constant region is a constant region of human
immunoglobulin. In some embodiments, the immunoglobulin constant
region is a constant region of human IgG. In some embodiments, the
antibody or antigen binding fragment of the present disclosure
comprises a heavy chain constant region of human IgG1, IgG2, IgG3,
IgG4, IgA1, IgA2 or IgM. In some embodiments, the antibody or
antigen binding fragment of the present disclosure comprises a
heavy chain constant region of human IgG1. In some embodiments, the
antibody or antigen binding fragment of the present disclosure
comprises a constant region of human immunoglobulin kappa 1 light
chain. In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises a constant region of
human immunoglobulin lambda light chain.
[0142] In some embodiments, the antibody or antigen binding
fragment of the present disclosure comprises one or more amino acid
residue substitutions or modifications yet retains specific binding
affinity to RBD of spike protein of SARS-CoV-2.
[0143] In some embodiments, the antibody or antigen binding
fragment is an affinity variant, a glycosylation variant, a
cysteine-engineered variant, or an Fc variant.
[0144] In some embodiments, the glycosylation variant comprises a
mutation at N297 (e.g. N297A, N297Q, or N297G), for example, to
modify the glycosylation site.
[0145] In some embodiments, the Fc variant comprises one or more
amino acid residue modifications or substitutions resulting in
increased effector functions relative to a wildtype Fc. In some
embodiments, the Fc variant comprises one or more amino acid
substitution(s) at one or more of the positions selected from the
group consisting of: 234, 235, 236, 238, 239, 240, 241, 243, 244,
245, 246, 247, 248, 249, 252, 254, 255, 256, 258, 260, 262, 263,
264, 265, 267, 268, 269, 270, 272, 274, 276, 278, 280, 283, 285,
286, 289, 290, 292, 293, 294, 295, 296, 298, 299, 300, 301, 303,
304, 305, 307, 309, 312, 313, 315, 320, 322, 324, 325, 326, 327,
329, 330, 331, 332, 333, 334, 335, 337, 338, 339, 340, 345, 360,
373, 376, 378, 382, 388, 389, 396, 398, 414, 416, 419, 430, 433,
434, 435, 436, 437, 438, 439 and 440 of the Fc region, wherein the
numbering of the residues in the Fc region is that of the EU index
as in Kabat. In some embodiments, the Fc variant comprises one or
more amino acid substitution selected from the group consisting of
234Y, 235Q, 236A, 236W, 239D, 239E, 239M, 243L, 2471, 267E, 268D,
268E, 268F, 270E, 280H, 290S, 292P, 298A, 298D, 298V, 300L, 3051,
324T, 326A, 326D, 326W, 330L, 330M, 333S, 332D, 332E, 333A, 334A,
334E, 339D, 339Q, 345R, 396L, 430G, 440Y, and any combination
thereof. In some embodiments, the Fc variant having increased
effector function comprises a combination of mutations selected
from the group consisting of: a) S239D, I332E, and A330L; b) F243L,
R292P, Y300L, V305I and P396L; c) S239D and I332E; d) S239D, I332E
and A330L; e) S298A, E333A and K334A; f) L234Y, L235Q, G236W,
S239M, H268D, D270E and S298A (in one heavy chain) and D270E,
K326D, A330M and K334E (in the opposing heavy chain); G236A, S239D
and I332E; g) K326W and E333S; h) S267E, H268F and S324T; i) E345R,
E430G and S440Y.
[0146] In some embodiments, the Fc variant comprises one or more
amino acid residue modifications or substitutions resulting in
reduced effector functions relative to a wildtype Fc. In some
embodiments, the Fc variant comprises one or more amino acid
substitution(s) at a position selected from the group consisting
of: 220, 226, 229, 233, 234, 235, 236, 237, 238, 267, 268, 269,
270, 297, 309, 318, 320, 322, 325, 328, 329, 330, and 331 of the Fc
region, wherein the numbering of the residues in the Fc region is
that of the EU index as in Kabat. In some embodiments, the Fc
variant comprises one or more amino acid substitution(s) selected
from the group consisting of 220S, 226S, 228P, 229S, 233P, 234V,
234G, 234A, 234F, 234A, 235A, 235G, 235E, 236E, 236R, 237A, 237K,
238S, 267R, 268A, 268Q, 269R, 297A, 297Q, 297G, 309L, 318A, 322A,
325L, 328R, 330S, 331S and any combination thereof. In some
embodiments, the Fc variant having reduced effector function
comprises a combination of mutations selected from the group
consisting of: a) K322A, L234A, and L235A; b) P331S, L234F, and
L235E; c) L234A and L235A; c) N297A; d) N297Q; e) N297G; f) L235E;
g) L234A and L235A (IgG1); h) F234A and L235A (IgG4); i) H268Q,
V309L, A330S and P331S (IgG2); j) V234A, G237A, P238S, H268A,
V309L, A330S and P331S (IgG2).
[0147] In some embodiments, the Fc variant comprises one or more
amino acid residue modifications or substitutions resulting in
improved binding affinity to neonatal Fc receptor (FcRn) at pH 6.0
while retaining minimal binding at pH 7.4, or increased serum half
life of the antibody. In some embodiments, the Fc variant comprises
one or more amino acid substitution(s) at a position selected from
the group consisting of: 234 (e.g., with F), 235 (e.g., with Q),
238 (e.g., with D), 250 (e.g., with E or Q), 252 (e.g., with
L/Y/F/W or T), 254 (e.g., with S or T), 256 (e.g., with S/R/Q/E/D
or T); 259 (e.g., with I); 272 (e.g., with A), 305(e.g., with A),
307(e.g., with A or P), 308 (e.g., with F, C or P), 311(e.g., with
A or R), 312 (e.g., with A), 322 (e.g., Q), 328 (e.g. E), 331
(e.g., with A), 378 (e.g., with A), 380 (e.g., with A), 382 (e.g.,
with A), 428 (e.g., with L or F), 432 (e.g., with C), 433 (e.g.,
with H/L/R/S/P/Q or K), 434 (e.g., with H/F or Y or S or A or W),
435 (e.g. with H), 436 (e.g., with L) and 437 (e.g., with C)) (all
positions by EU numbering). In some embodiments, the Fc variant
comprises one or more amino acid substitution(s) selected from the
group consisting of 234F, 235Q, 238D, 250Q, 252T, 252Y, 254T, 256E,
2591, 272A, 305A, 307A, 308F, 311A, 322Q, 328E, 331S, 380A, 428L,
432C, 433K, 433S, 434S, 434Y, 434F, 434W, 434A, 435H, 436L, 437C
and any combination thereof. In some embodiments, the Fc variant
having increased serum half-life or improved pH-dependent binding
to FcRn comprises a combination of mutations selected from the
group consisting of: a) M428L and N434S; b) P238D and L328E; c)
M252Y, S254T and T256E; d) L234F, L235Q, K322Q, M252T, S254T and
T256E; e) M428L, V259I and V308F; f) H433K and N434Y; g) H433K and
N434F; h) T250Q and M428L; i) T307A, E380A and N434A; and j) 432C,
433S, 434W, 435H, 436L, 437C.
[0148] In some embodiments, at least one of the substitutions or
modifications is in one or more of the CDR sequences. In some
embodiments, at least one of the substitutions or modifications is
in one or more of the non-CDR sequences of the heavy chain variable
region or light chain variable region. In some embodiments, at
least one of the substitutions is a conservative substitution.
[0149] In some embodiments, the antibody or antigen binding
fragment of the present disclosure is a monoclonal antibody, a
bispecific antibody, a multi-specific antibody, a recombinant
antibody, a labeled antibody, a bivalent antibody, an
anti-idiotypic antibody, a fusion protein, or a dimerized or
polymerized antibody, or a modified antibody (e.g. glycosylated
antibody). In some embodiments, the antibody or antigen binding
fragment of the present disclosure is a diabody, a Fab, a Fab', a
F(ab').sub.2, a Fd, an Fv fragment, a disulfide stabilized Fv
fragment (dsFv), a (dsFv).sub.2, a bispecific dsFv (dsFv-dsFv'), a
disulfide stabilized diabody (ds diabody), a single-chain antibody
molecule (scFv), an scFv dimer (bivalent diabody), a bispecific
scFv dimer, a multispecific antibody, a heavy chain antibody, a
camelized single domain antibody, a nanobody, a domain antibody, or
a bivalent domain antibody. In some embodiments, the antibody or
antigen binding fragment of the present disclosure is a full human
antibody.
[0150] In some embodiments, the antibody or antigen binding
fragment of the present disclosure is linked to one or more
conjugate moieties. In some embodiments, the conjugate moiety
comprises a therapeutic agent, a radioactive isotope, a detectable
label, a pharmacokinetic modifying moiety, or a purifying moiety.
In some embodiments, the conjugate moiety is covalently attached
either directly or via a linker.
[0151] In one aspect, the present disclosure provides an isolated
or recombinant antibody or an antigen-binding fragment thereof,
which competes for binding to RBD of spike protein of SARS-CoV-2
with the antibody or an antigen-binding fragment thereof described
herein.
[0152] In another aspect, the present disclosure provides
bispecific antibody molecules comprising an anti-SARS-CoV-2
antibody or antigen-binding fragment thereof as disclosed
herein.
[0153] In certain embodiments, the bispecific or bivalent
antibodies provided herein comprises a first antigen-binding domain
and a second antigen-binding domain, wherein the first
antigen-binding domains is derived from a monoclonal antibody
selected from the group consisting of P2A-1A8, P2A-1A9, P2B-2G11,
P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4, P2C-1A3, P2C- 1C8, P2C-1C10,
P2C-1D5, P2C-1F11, P2B-1G5, P2B-1A1, P2C-1D7, P2B-1A10, P2B-1D9,
P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2, P5A-2F11,
P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6, P5A-1B8,
P5A-1B9, P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9, P5A-2H3, P5A-3A1,
P5A-3A6, P5A-3B4, P5A-3C12, and P22A-1D1. The second
antigen-binding domain can be derived from any suitable
antibody.
[0154] In certain embodiments, the bispecific antibodies provided
herein comprises a first antigen-binding domain and a second
antigen-binding domain, wherein the first and the second
antigen-binding domains are derived from any two monoclonal
antibodies selected from the group consisting of P2A-1A8, P2A-1A9,
P2B-2G11, P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4, P2C-1A3, P2C-1C8,
P2C-1C10, P2C-1D5, P2C-1F11, P2B-1G5, P2B-1A1, P2C-1D7, P2B-1A10,
P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2,
P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6,
P5A-1B8, P5A-1B9, P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9, P5A-2H3,
P5A-3A1, P5A-3A6, P5A-3B4, P5A- 3C12, and P22A-1D1. In certain
embodiments, the first and the second antigen-binding domains are
derived from P2C-1F11 and P2B-2F6, respectively. In certain
embodiments, the first and the second antigen-binding domains are
derived from P2C-1F11 and P2B-1G5, respectively.
[0155] In certain embodiments, the bispecific antibody molecules
have at least two distinct antigen-binding sites with different
specificities.
[0156] In certain embodiments, the bispecific antibody molecules
provided herein are capable of binding to different epitopes on the
spike protein of SARS-CoV-2 virus. In some embodiments, the two or
more antibodies bind to different epitopes in RBD of spike protein
of SARS-CoV-2.
[0157] In certain embodiments, the bispecific antibody molecules
provided herein has a first antigen-binding domains specificity
directed to the RBD of the spike protein of SARS-CoV-2 virus and a
second antigen-binding domains specificity directed to a second
antigen.
[0158] In another aspect, the present disclosure provides an
isolated polynucleotide encoding the antibody or antigen binding
fragment thereof as described herein.
[0159] In some embodiments, the isolated polynucleotide of the
present disclosure comprises a nucleotide sequence selected from a
group consisting of: SEQ ID NOs: 9-10, 19-20, 29-30, 39-40, 49-50,
59-60, 63-64, 73-74, 83-84, 93-94, 103-104, 113-114, 144-145,
154-155, 164-165, 174-175, 184-185, 194-195, 204-205, 214-215,
224-225, 234-235, 244-245, 254-255, 264-265, 274-275, 284-285,
294-295, 304-305, 314-315, 324-325, 334-335, 344-345, 354-355,
364-365, 374-375, 384-385, 394-395, 404-405, 414-415, 424-425, and
434-435, or a homologous sequence thereof having at least 80%
sequence identity.
[0160] In some embodiments, the homologue sequence encodes the same
protein as encoded by any nucleotide sequence selected from the
group consisting of SEQ ID NOs: 9-10, 19-20, 29-30, 39-40, 49-50,
59-60, 63-64, 73-74, 83-84, 93-94, 103-104, 113-114, 144-145,
154-155, 164-165, 174-175, 184-185, 194-195, 204-205, 214-215,
224-225, 234-235, 244-245, 254-255, 264-265, 274-275, 284-285,
294-295, 304-305, 314-315, 324-325, 334-335, 344-345, 354-355,
364-365, 374-375, 384-385, 394-395, 404-405, 414-415, 424-425, and
434-435.
[0161] In one aspect, the present disclosure provides a vector
comprising the isolated polynucleotide of the present disclosure.
In some embodiments, said vector is an expression vector.
[0162] In one aspect, the present disclosure provides a host cell
comprising the vector of the present disclosure.
[0163] In one aspect, the present disclosure provides a method of
producing the antibody or antigen binding fragment of the present
disclosure. In some embodiments, the method comprises culturing the
host cell of the present disclosure under the condition at which
the expression vector of the present disclosure is expressed. In
some embodiments, the method of the present disclosure further
comprises purifying the antibody produced by the host cell.
[0164] In some embodiments, the pharmaceutical composition
disclosed herein can comprise a combination of two or more
antibodies or antigen binding fragments of the present disclosure.
In some embodiments, the pharmaceutical composition comprises a
combination of two or more monoclonal antibodies, each of which
comprises heavy chain CDR sequences and light chain CDR sequences
derived from an antibody selected from the group consisting of
P2A-1A8, P2A-1A9, P2B-2G11, P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4,
P2C-1A3, P2C-1C8, P2C-1C10, P2C-1D5, P2C-1F11, P2B-1G5, P2B-1A1,
P2C-1D7, P2B-1A10, P2B-1D9, P2B- 1E4, P2B-1G1, P4A-2D9, P5A-2G7,
P5A-3C8, P5A-1D2, P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6,
P4B-1F4, P5A-1B6, P5A-1B8, P5A-1B9, P5A-1D1, P5A-1D10, P5A- 2D11,
P5A-2G9, P5A-2H3, P5A-3A1, P5A-3A6, P5A-3B4, P5A-3C12, and
P22A-1D1.
[0165] In certain embodiments, the pharmaceutical composition
comprises a first antibody comprising heavy chain CDR sequences and
light chain CDR sequences derived from P2C-1F11, and a second
antibody comprising heavy chain CDR sequences and light chain CDR
sequences derived from antibody P2B-2F6. In certain embodiments,
the pharmaceutical composition comprises a first antibody
comprising heavy chain CDR sequences and light chain CDR sequences
derived from P2C-1F11, and a second antibody comprising heavy chain
CDR sequences and light chain CDR sequences derived from antibody
P2B-1G5.
[0166] In some embodiments, the two or more antibodies or antigen
binding fragments bind to different epitopes in RBD of spike
protein of SARS-CoV-2. In some embodiments, the two or more
antibodies comprise a first antibody which comprises P2C-1F11 or an
antigen binding fragment thereof, and a second antibody which is
selected from the group consisting of P2C-1A3, P2C-1C10, P2B-2F6,
P2B-1G5, and P2A-1B3, or an antigen binding fragment thereof. In
some embodiments, the two or more antibodies comprise a first
antibody which comprises P2C-1A3 or an antigen binding fragment
thereof, and a second antibody which is selected from the group
consisting of P5A-3C8, P5A-1D2, P22A-1D1, P2C-1F11, and P2A-1B3, or
an antigen binding fragment thereof. In some embodiments, the two
or more antibodies comprise a first antibody which comprises
P2B-2F6 or an antigen binding fragment thereof, and a second
antibody selected from the group consisting of P5A-3C8, P5A-1D2,
P22A-1D1, P2C-1C10, P2C-1F11, P2B-1G5, and P2A-1B3, or an antigen
binding fragment thereof. In some embodiments, the two or more
antibodies comprise a first antibody which comprises P2A-1B3 or an
antigen binding fragment thereof, and a second antibody selected
from the group consisting of P5A-3C8, P5A-1D2, P22A-1D1, P2C-1A3,
P2C-1C10, P2C-1F11, P2B-2F6, and P2A-1A10, or an antigen binding
fragment thereof. In some embodiments, the two or more antibodies
comprise a first antibody which comprises P2C-1C10 or an antigen
binding fragment thereof, and a second antibody selected from the
group consisting of P5A-3C8, P5A-1D2, P22A-1D1, P2C-1A3, P2C-1F11,
and P2A-1B3, or an antigen binding fragment thereof
[0167] In some embodiments, the pharmaceutical compositions
comprise the polynucleotides encoding the anti-SARS-CoV-2
antibodies or the antigen-binding fragments thereof, and one or
more pharmaceutically acceptable carriers. The present disclosure
further provides pharmaceutical compositions comprising the
polynucleotides encoding the combination of the two or more
anti-SARS-CoV-2 antibodies or the antigen-binding fragments
thereof, and one or more pharmaceutically acceptable carriers. In
certain embodiments, the polynucleotides comprise an expression
vector. In certain embodiments, the expression vector comprises a
viral vector or a non-viral vector. In certain embodiments, the
expression vector is suitable for gene therapy in human. In certain
embodiments, the expression vector comprises a DNA vector or a RNA
vector.
[0168] In some embodiments, the pharmaceutical composition further
comprises a second bioactive agent, such as a second therapeutic
agent or a second prophylactic agent.
[0169] In one aspect, the present disclosure provides a kit for
detecting a SARS-CoV-2 antigen, comprising the antibody or antigen
binding fragment of the present disclosure. In some embodiments,
the kit of further comprises a control reagent comprising RBD of
spike protein of the SARS-CoV-2. In some embodiments, the kit
further comprises a set of reagents for detecting complex of the
antibody or the antigen-binding fragment bound to the SARS-CoV-2
antigen.
[0170] In one aspect, the present disclosure provides a method of
treating SARS-CoV-2 infection in a subject. The present disclosure
also provides methods of treating a disease, disorder or condition
associated with SARs-CoV-2 infection in a subject. In some
embodiments, the method comprises administering a therapeutically
effective amount of one or more of the antibody, the antigen
binding fragment, or one or more polynucleotides encoding one or
more of the antibody or antigen-binding fragment thereof provided
herein, or the pharmaceutical composition of the present disclosure
to the subject.
[0171] In one aspect, the present disclosure provides a method of
preventing SARS-CoV-2 infection in a subject. The present
disclosure also provides methods of preventing a disease, disorder
or condition associated with SARs-CoV-2 infection in a subject. In
some embodiments, the method comprises administering a
prophylactically effective amount of one or more of the antibody or
antigen binding fragment, or the pharmaceutical composition of the
present disclosure to the subject.
[0172] In some embodiments, the administration is via oral, nasal,
intravenous, subcutaneous, or intramuscular administration. In some
embodiments, the subject is human. In some embodiments, the
polynucleotide provided herein can be administered to a subject by,
for example, transfection techniques such as electroporation, or
hydrodynamic injection. In some embodiments, the polynucleotides
comprise viral vectors such as AAV, and can be administered via
local injection (e.g. intramuscular, intranasal, intradermal,
subcutaneous, etc.) or systematic administration (e.g. intravenous
administration).
[0173] In some embodiments, the method further comprises
administering a therapeutically effective amount of a second
bioactive agent which can be a therapeutic agent or a prophylactic
agent. In some embodiments, the second therapeutic agent is an
anti-viral agent. In some embodiments, an anti-viral agent
comprises an antiviral peptide, an anti-viral antibody, an
anti-viral compound, an anti-viral cytokine, or an anti-viral
oligonucleotide. In some embodiments, the second therapeutic agent
is an RNA dependent RNA polymerase inhibitor, a non-nucleoside
reverse transcriptase inhibitor (NNRTI), nucleoside reverse
transcriptase inhibitor (NRTI), purine nucleoside, antiviral
interferon, adamantine antiviral compound, or any other suitable
antiviral agent. In some embodiments, the second therapeutic agent
is remdesivir, chloroquine, hydroxychloroquine, lopinavir,
ritonavir, APN01, favilavir, mesalazine, toremifene, eplerenone,
paroxetine, sirolimus, dactinomycin, irbesartan, emodin,
mercaptopurine, melatonin, quinacrine, carvedilol, colchicine,
camphor, equilin, oxymetholone, nafamosta, camostat, baricitinib,
darunavir, ribavirin, galidesivir, BCX-4430, Arbidol, nitazoxanide,
derivatives thereof, or any combination thereof.
[0174] In one aspect, the present disclosure provides a method of
detecting presence or amount of SARS-CoV-2 virus antigen in a
sample. In some embodiments, the method comprises contacting the
sample with one or more of the antibody or antigen binding fragment
of the present disclosure, and determining the presence or the
amount of the SARS-CoV-2 virus antigen in the sample.
[0175] In one aspect, the present disclosure provides use of one or
more of the antibody or antigen binding fragment of the present
disclosure in the manufacture of a medicament for treating or
preventing SARS-CoV-2 infection or a disease, disorder or condition
associated with SARs-CoV-2 infection. In one aspect, the present
disclosure provides use of one or more of the antibody or antigen
binding fragment of the present disclosure in the manufacture of a
medicament for preventing, managing, treating and/or ameliorating
in a subject a disease or a disorder caused by or associated with
coronavirus (e.g. SARs-COV-2) infection and/or a symptom or
respiratory condition relating thereto.
[0176] In one aspect, the present disclosure provides use of one or
more of the antibody or antigen binding fragment of the present
disclosure in the manufacture of a diagnostic reagent for detecting
SARS-CoV-2 infection.
[0177] In one aspect, the present disclosure provides a kit for
detecting an antibody capable of specifically binding to
receptor-binding domain (RBD) of the spike protein of SARS-CoV-2,
comprising a polypeptide comprising an amino acid sequence
comprising SEQ ID NO: 128. In some embodiments, the polypeptide is
immobilized on a substrate. In some embodiments, the kit further
comprises a set of reagents for detecting complex of the antibody
bound to the polypeptide.
[0178] In one aspect, the present disclosure provides a method of
detecting presence or amount of an antibody capable of specifically
binding to RBD of the spike protein of SARS-CoV-2 in a sample,
comprising contacting the sample with a polypeptide comprising an
amino acid sequence comprising SEQ ID NO: 128, and determining the
presence or the level of the antibody in the sample. In some
embodiments, the absence of the antibody in the sample or the level
of the antibody in the sample being below a threshold indicates
that the subject is more likely to suffer from disease
progression.
[0179] In another aspect, the present disclosure provides a method
of determining the likelihood of disease progression in a subject
infected with SARS-CoV-2, the method comprising: contacting a
sample obtained from the subject with a polypeptide comprising an
amino acid sequence comprising SEQ ID NO: 128, and detecting the
presence or the level of an antibody in the sample wherein the
antibody is capable of specifically binding to RBD of the spike
protein of the SARS-CoV-2, wherein the subject is likely to
experience disease progression when the antibody in the sample is
absent or is below a threshold.
[0180] In yet another aspect, the present disclosure provides a
method of monitoring treatment response in a subject infected with
SARS-CoV-2 and received a treatment, the method comprising: (i)
contacting a sample from the subject with a peptide comprising an
amino acid sequence of SEQ ID NO: 128; (ii) detecting a first level
of an antibody in the sample wherein the antibody is capable of
specifically binding to RBD of the spike protein of the SARS-CoV-2;
and (iii) comparing the first level of the antibody with a second
level of the antibody detected in the subject prior to the
treatment; wherein the first level being higher than the second
level indicates that the subject is responsive to the
treatment.
[0181] In yet another aspect, the present disclosure provides a
method of neutralizing SARS-CoV-2 in a subject or in a sample in
vitro, comprising administering a therapeutically effective amount
of one or more of the antibody or antigen binding fragment thereof
provided herein, or the pharmaceutical composition provided herein
to the subject or to the sample.
[0182] In yet another aspect, the present disclosure provides a
crystal of RBD of the spike protein of SARS-CoV-2 in complex with
an antibody. In some embodiments, the antibody in complex with the
RBD comprises a heavy chain variable region of SEQ ID NO: 47 and a
light chain variable region of SEQ ID NO: 48. In some embodiments,
the antibody in complex with the RBD comprises a heavy chain
variable region of SEQ ID NO: 111 and a light chain variable region
of SEQ ID NO: 112.
[0183] In some embodiment, the crystal has or consists of a
P2.sub.12.sub.12.sub.1 space group with unit cell dimensions of
a=70.23 .ANG., b=90.15 .ANG., and c=112.35 .ANG..
[0184] In some embodiment, the crystal has or consists of a C121
space group with unit cell dimensions of a=194.88 .ANG., b=85.39
.ANG., and c=58.51 .ANG..
[0185] In some embodiment, the crystal has or consists of a C2
space group with unit cell dimensions of a=193.34 .ANG., b=86.60
.ANG., and c=57.16 .ANG..
[0186] In some embodiment, the crystal has or consists of a C2
space group with unit cell dimensions of a=158.75 .ANG., b=67.51
.ANG., and c=154.37 .ANG..
[0187] In some embodiment, the crystal has or consists of a
P2.sub.12.sub.12.sub.1 space group with unit cell dimensions of
a=112.54 .ANG., b=171.57 .ANG., and c=54.87 .ANG..
BRIEF DESCRIPTION OF FIGURES
[0188] The following drawings form part of the present
specification and are included to further demonstrate certain
aspects of the present disclosure. The disclosure may be better
understood by reference to one or more of these drawings in
combination with the detailed description of specific embodiments
presented herein.
[0189] FIG. 1A-FIG. 1F. Analyses of plasma and B cell responses
specific to SARS-CoV-2. Serial dilutions of plasma samples were
analyzed for binding to the (FIG. 1A) RBDs or (FIG. 1B) trimeric
Spikes of SARS-CoV-2, SARS-CoV and MERS-CoV by ELISA and (FIG. 1C)
for neutralizing activity against pseudoviruses bearing envelope
glycoprotein of SARS-CoV-2, SARS-CoV and MERS-CoV. Binding to
SARS-CoV-2 NP protein was also evaluated (A). All results were
derived from at least two independent experiments. (FIG. 1D) Gating
strategy for analysis and isolation of RBD-specific memory B cells
and (FIG. 1E and FIG. 1F) their representation among the total and
memory subpopulation of B cells in the eight study subjects.
Samples were named as either A, B, or C depending on collection
sequence. FSC-W, forward scatter width; FSC-A, forward scatter
area; and SSC-A side scatter area.
[0190] FIG. 2. Heavy chain repertoires of SARS-CoV-2 RBD-specific
antibodies analyzed by individual subject. Distribution and
frequency of heavy chain variable (VH) genes usage in each subject
shown along the horizontal bar. The same color scheme is used for
each VH family across all study subjects. The VHs that dominate
across isolated antibodies are indicated by actual frequencies in
their respective color boxes. The number of RBD-binding antibodies
versus total antibodies isolated are shown on the right.
[0191] FIG. 3A-FIG. 3F. Clonal expansion of specific heavy and
light chain families in the P#2 antibody repertoire. (FIG. 3A)
Phylogenetic analysis of VH (left) and VL 20 (right) genes for all
RBD-binding antibodies. Clonal expanded VH and VL clusters are
paired and highlighted in three different colors. Branch lengths
are drawn to scale so that sequence relatedness can be readily
assessed. (FIG. 3B)-(FIG. 3C) Clonal expansion in relation to
members of other VH and VL families based on somatic hypermutations
(SHM) and CDR3 loop lengths. For the pie charts of VH (left) and VL
(right) genes, the radii represent the CDR3 loop length and the
color scale indicates the degree of SHM. Heavy and light chain
repertoires for each antibody are shown along the pie circles.
(FIG. 3D)-(FIG. 3E) Lineage analysis for heavy and light chains in
pie charts. The numbers in the center represent the number of
RBD-specific antibodies. Each slice represents a unique clone and
proportional to its own size. (FIG. 3F) Counts of various HCDR3
length from IGHV3-53 and IGHV3-66 as well as RBD binders.
[0192] FIG. 4A-FIG. 4V. Antibody binding, competition with ACE2,
and neutralization analyzed by pseudovirus and live SARS-CoV-2.
(FIG. 4A) Binding kinetics of representative mAbs to SARS-CoV-2 RBD
measured by SPR. The black lines indicate the experimentally
derived curves while the grey lines represent fitted curves based
on the experimental data. (FIG. 4B) Antibody and ACE2 competition
for binding to SARS-CoV-2 RBD measured by SPR. The sensorgrams show
distinct binding patterns of ACE2 to SARS-CoV-2 RBD with or without
prior incubation with each representative antibody. (FIG. 4C and
FIG. 4D) Antibody neutralization analyzed by SARS-CoV-2 RBD binding
assay. (FIG. 4E through FIG. 4R) Antibody neutralization analyzed
by pseudovirus assay. (FIG. 4S)-(FIG. 4T) Antibody neutralization
analyzed by live SARS-CoV-2 neutralization assay, in which dashed
lines indicated 50% reduction in viral infectivity. VRC01 is an
HIV-1 specific antibody and used here as a negative control. (FIG.
4U)-(FIG. 4V) Summary of actual values from studies in FIG. 4A
through FIG. 4T. Antibody binding to RBD was presented either by Kd
or by competing with ACE2 where "+++" indicates >80%
competition; "++" indicates 50-80%; "+" indicates 20-50%; and "-"
indicates<20%. IC.sub.50 represents the half-maximal whereas
IC.sub.80 the 80% inhibitory concentrations and IC.sub.90 the 90%
inhibitory concentrations tested in the pseudovirus and live
SARS-CoV-2 neutralization assay. Only the antibody heavy chains are
indicated at the upper left corner for their family designation,
CDR3 length, and SHM in relative to corresponding germline ancestor
sequence. n.d. not done.
[0193] FIG. 5A-FIG. 5T. Crystal structures of 2F6 and P2C-1F11 in
complex with SARS-CoV-2 RBD respectively, and the lists of
determined contacting residues at the antibody/SARS-CoV-2
interfaces. (FIG. 5A) Overall structure of 2F6 Fab in complex with
SARS-CoV-2 RBD. (FIG. 5B) The critical interactions between 2F6 and
SARS-CoV-2 RBD. (FIG. 5C) 2F6/RBD complex was aligned to ACE2/RBD
complex (PDB ID: 6M0J). Circle indicated the clash between ACE2 and
2F6. (FIG. 5D) The SARS-CoV-2 spike (PDB ID: 6VSB) is shown as a
molecular surface, with each protomer colored either light orange,
blue and green. 2F6/RBD complex could be aligned to both the "up"
RBD (light orange) and the "down" RBD (light blue) in spike. 2F6
heavy chain is colored in magenta, 2F6 light chain in yellow,
SARS-CoV-2 RBD in cyan, and ACE2 in green. (FIG. 5E) Contacting
residues at the SARS-CoV-2/2F6 interface. (FIG. 5F) Overall
structure of P2C-1F11 Fab in complex with SARS-CoV-2 RBD. (FIG. 5G)
Contacting residues at the SARS-CoV-2/1F11 interface. (FIG.
5H)-(FIG. 5K) Overall structures, crystal structures of the RBD and
Fab complexes and RBD binding residues shared with ACE2 were shown
for antibodies P22A-1D1, P5A-1D2, P5A-3C8 and P2C-1F11
respectively. (FIG. 5I) The footprint of Fabs and ACE2 on
SARS-CoV-2 RBD. The color of the epitope was depicted as in panel
(FIG. 5H). The epitope of ACE2 was colored by green. (J) (FIG. 5M)
Conserved HCDR1, HCDR2 and different HCDR3. RBD was shown as
surface. CDR loops of the heavy chain were shown as ribbons. (FIG.
5N) The interactions between the three conserved tyrosine at HCDR1
and HCDR2. (FIG. 5O) The interactions between HCDR2 -SGGS- segment
and RBD. Hydrogen bonds were shown as black dashed line and
P5A-3C8/RBD complex was used as an example in panel L and M. Y505
residue on RBD protruded into the wedge hole of the light chain For
P22A-1D1 (FIG. 5Q), P5A-3C8 (FIG. 5R) and P2C-1F11 (FIG. 5S),
whereas for P5A-1D2 (FIG. 5P) Y505 displayed a different
conformation because of the binding of the long HCDR3. (FIG. 5T)
Summary of contacts between SARS-CoV-2 RBD and P22A-1D1, P5A-3C8,
P5A-1D2, P2C-1F11 (distance cutoff 4 .ANG.).
[0194] FIG. 6A-FIG. 6C. Analysis of plasma binding to cell surface
expressed trimeric Spike protein. (FIG. 6A) and (FIG. 6B) HEK 293T
cells transfected with expression plasmid encoding the full length
spike of SARS-CoV-2, SARS-CoV or MERS-CoV were incubated with 1:100
dilutions of convalescent plasma from the study subjects. The cells
were then stained with PE labeled anti-human IgG Fc secondary
antibody and analyzed by FACS. Positive control antibodies include
S230 and m396 targeting the RBD of SARS-CoV Spike, and Mab-GD33
targeting the RBD of MERS-CoV Spike. VRC01 is negative control
antibody targeting HIV-1 envelope glycoprotein. (FIG. 6C) Impact of
epitope mutations on binding of antibodies P22A-1D1, P5A-1D2,
P5A-3C8, P2C-1F11 and P2B-2F6. Cell surface expressed wild type or
mutant spike glycoprotein of SARS-Cov-2 were incubated with the
ACE2, the tested antibodies, and control antibodies S2 mAb,
followed by staining with anti-human IgG Fc PE (for identified
human antibodies), anti-mouse IgG FITC (for S2 mAb) or anti-his PE
(for ACE2) secondary antibody and analyzed by FACS. P2B-2F6
recognizes a distinct epitope on SARS-CoV-2 RBD from those public
antibodies and used here as positive control for the S1 protein of
the spike. S2 is a mouse monoclonal specific for S2 protein of the
spike. The cell percentage in the gate are shown. Mutants that
resulted in more than 80% reduction in binding are highlighted in
either grey boxes for public antibodies or in orange boxes for
ACE2. The percent reduction was determined by the MFI weighted by
multiplying the number of positive cells in the selected gates and
normalized in relative to that of wild type and S2 control. Data
shown were from at least two independent experiments.
[0195] FIG. 7A-FIG. 7K. RBD-specific memory B cells analyzed and
isolated through FACS (FIG. 7A)-(FIG. 7K). The recombinant RBD was
labeled with either a Strep or His tag and used alone or in
combination to identify and isolate RBD-specific single B cells
through staining with the Streptavidin-APC and/or Streptavidin-PE,
or anti-His-APC and anti-His-PE antibodies. B cells to be isolated
are highlighted in boxes or ovals. Samples were named as either A,
B, or C depending on collection sequence. FSC-W, forward scatter
width; FSC-A, forward scatter area; and SSC-A side scatter
area.
[0196] FIG. 8. ELISA screening of SARS-CoV-2 RBD-specific
antibodies in the supernatant of transfected cells. The study
subjects and the date of sampling are indicated on the top. Samples
were named as either A, B, or C depending on collection sequence.
Antibodies tested for each sample are aligned in one vertical
column whenever possible. For each evaluated antibody, at least two
independent measurements were performed and are presented
adjacently on the same row. Binding activities were assessed by OD
450 and indicated by the color scheme on the right. Negatives (no
binding activity) are shown in gray for OD 450 values less than
0.1.
[0197] FIG. 9A-FIG. 9G. Binding kinetics of isolated mAbs with
SARS-CoV-2 RBD measured by SPR and ELISA respectively. (FIG. 9A,
FIG. 9B) For SPR, the purified soluble SARS-CoV-2 RBD, SARS-CoV RBD
and MERS-CoV2 RBD were covalently immobilized onto a CM5 sensor
chip followed by injection of individual antibody at five different
concentrations. The black lines indicate the experimentally derived
curves while the grey lines represent fitted curves based on the
SPR experimental data. For ELISA analysis, recombinant SARS-CoV RBD
and MERS-CoV2 RBD were coated on the ELISA plate, and a serial
dilution of each antibody was evaluated against SARS-CoV RBD and
MERS-CoV2 RBD coated plates respectively and their binding activity
was recorded at an optical density (OD) of 450 nm and 630 nm. (FIG.
9C-FIG. 9F) For ELISA analysis, S230 was used as a positive control
antibody against SARS-COV, MAB-GD33 was used as a positive control
antibody against MERS-COV, and VRC01 was used as negative control
antibody. (FIG. 9G) Binding kinetics of isolated mAbs with
SARS-CoV-2 RBD were measured by SPR.
[0198] FIG. 10A-FIG. 10C. Antibody and ACE2 competition for binding
to SARS-CoV-2 RBD measured by SPR for patient #2 (FIG. 10A),
patient #1 (FIG. 10B), and patient#5, #22 and #2 (FIG. 10C). The
sensorgrams show distinct binding patterns of ACE2 to SARS-CoV-2
RBD with (corresponding to curves for "antibody+ACE2") or without
(corresponding to curves for "ACE2") prior incubation with each
testing antibody. The competition capacity of each antibody is
indicated by the level of reduction in response unit of ACE2
comparing with or without prior antibody incubation.
[0199] FIG. 11A-FIG. 11B. Analysis of antibody binding to cell
surface expressed trimeric Spike protein. HEK 293T cells
transfected with expression plasmid encoding the full length spike
of SARS-CoV-2, SARS-CoV or MERS-CoV were incubated with 20 ug/ml
testing antibodies (FIG. 11A) and (FIG. 11B). The cells were then
stained with PE labeled anti-human IgG Fc secondary antibody and
analyzed by FACS. Positive control antibodies include S230 and m396
targeting the RBD of SARS-CoV Spike, and Mab-GD33 targeting the RBD
of MERS-CoV Spike. VRC01 is the negative control antibody targeting
HIV-1 envelope glycoprotein.
[0200] FIG. 12A-FIG. 12D. Neutralization activity of mAbs against
live SARS-CoV-2 analyzed by focus reduction neutralization test
(FRNT). Serial dilution of each antibody was tested against live
SARS-CoV-2 infection. Their neutralizing activities are represented
by the reduction in the number of SARS-CoV-2 foci calculated by an
EliSpot reader (Cellular Technology Ltd) (FIG. 12A)-(FIG. 12D).
[0201] FIG. 13. Epitope mapping through competitive binding
measured by SPR. The sensorgrams show distinct binding patterns
when pairs of testing antibodies were sequentially applied to the
purified SARS-CoV-2 RBD covalently immobilized onto a CM5 sensor
chip. The level of reduction in response unit comparing with or
without prior antibody incubation is the key criteria for
determining the two mAbs recognize the separate or closely situated
epitopes.
[0202] FIG. 14. Multiple sequence alignment of the CDR1-CDR3
regions of the heavy chain sequences from the public clonetypes.
Included are antibodies P22A-1D1 (SEQ ID NO: 432), P5A-1D2 (SEQ ID
NO: 242), P5A-3C8 (SEQ ID NO: 232) and P2C-1F11(SEQ ID NO: 111)
along with IGVH3-53 (SEQ ID NO: 436) and/IGVH3-66(SEQ ID NO: 437),
a top germline allele assignment for public antibodies shown. Grey
letters show mutations from germline.
[0203] FIG. 15A-FIG. 15B. Antibody individual PK profiles of single
administration fit the population PK prediction model and the
measurement data for mAb1 (FIG. 15A) and mAb2 (FIG. 15B). Solid
lines: predicted medians. Dashed lines and the dots: measured
concentrations in the subjects. The shaded areas represent the
5th-95th percentiles.
[0204] FIG. 16. In vitro Neutralization Activity of mAb1 and mAb2
Combination in SARS-CoV-2 Live Virus Micro-Neutralization
Assay.
[0205] FIG. 17A-FIG. 17B. Mean serum concentration profiles of in
cynomolgus monkeys following single IV infusion administration of
at 10 mg/kg for mAb1 (FIG. 17A) and mAb2 (FIG. 17B).
DETAILED DESCRIPTION
[0206] Throughout the present disclosure, the articles "a", "an"
and "the" are used herein to refer to one or to more than one
(i.e., to at least one) of the grammatical object of the article.
By way of example, "an antibody" means one antibody or more than
one antibody.
[0207] The features and advantages of the disclosed compositions
and methods will be more readily understood, by those of ordinary
skill in the art, from reading the following detailed description.
It is to be appreciated that certain features of the disclosed
compositions and methods, which are, for clarity, described above
and below in the context of separate embodiments, may also be
provided in combination in a single embodiment. Conversely, various
features of the disclosed compositions and methods that are, for
brevity, described in the context of a single embodiment, may also
be provided separately or in any sub-combination. In addition,
references in the singular may also include the plural (for
example, "a" and "an" may refer to one, or one or more) unless the
context specifically states otherwise.
[0208] The use of numerical values in the various ranges specified
in this application, unless expressly indicated otherwise, are
stated as approximations as though the minimum and maximum values
within the stated ranges were both proceeded by the word "about."
In this manner, slight variations above and below the stated ranges
can be used to achieve substantially the same results as values
within the ranges. Also, the disclosure of these ranges is intended
as a continuous range including every value between the minimum and
maximum values.
[0209] The term "about" and its grammatical equivalents in relation
to a reference numerical value and its grammatical equivalents as
used herein can include a range of values plus or minus 10% from
that value, such as a range of values plus or minus 10%, 9%, 8%,
7%, 6%, 5%, 4%, 3%, 2%, or 1% from that value. For example, the
amount "about 10" includes amounts from 9 to 11.
[0210] Definitions
[0211] Antibody Related Terms
[0212] The term "antibody" as used herein includes any
immunoglobulin, monoclonal antibody, polyclonal antibody,
monovalent antibody, bivalent antibody, multivalent antibody,
bispecific antibody, multi-specific antibody that binds to a
specific antigen. A native intact antibody comprises two heavy (H)
chains and two light (L) chains. Mammalian heavy chains are
classified as alpha, delta, epsilon, gamma, and mu, each heavy
chain consists of a variable region (VH) and a first, second,
third, and optionally fourth constant region (CH1, CH2, CH3, CH4
respectively); mammalian light chains are classified as .lamda. or
.kappa., while each light chain consists of a variable region (VL)
and a constant region. The antibody has a "Y" shape, with the stem
of the Y consisting of the second and third constant regions of two
heavy chains bound together via disulfide bonding. Each arm of the
Y includes the variable region and first constant region of a
single heavy chain bound to the variable and constant regions of a
single light chain. The variable regions of the light and heavy
chains are responsible for antigen binding. The variable regions in
both chains generally contain three highly variable loops called
the complementarity determining regions (CDRs) (light chain CDRs
including LCDR1, LCDR2, and LCDR3, heavy chain CDRs including
HCDR1, HCDR2, HCDR3). CDR boundaries for the antibodies and
antigen-binding fragments disclosed herein may be defined or
identified by the conventions of Kabat, IMGT, Chothia, or
Al-Lazikani (Al-Lazikani, B., Chothia, C., Lesk, A. M., J Mol.
Biol., 273(4), 927 (1997); Chothia, C. et al., J Mol Biol. December
5; 186(3):651-63 (1985); Chothia, C. and Lesk, A. M., J Mol. Biol.,
196,901 (1987); Chothia, C. et al., Nature. December 21-28;
342(6252):877-83 (1989); Kabat E. A. et al., Sequences of Proteins
of immunological Interest, 5th Ed. Public Health Service, National
Institutes of Health, Bethesda, Md. (1991); Marie-Paule Lefranc et
al., Developmental and Comparative Immunology, 27: 55-77 (2003);
Marie-Paule Lefranc et al., Immunome Research, 1(3), (2005);
Marie-Paule Lefranc, Molecular Biology of B cells (second edition),
chapter 26, 481-514, (2015)). The three CDRs are interposed between
flanking stretches known as framework regions (FRs) (light chain
FRs including LFR1, LFR2, LFR3, and LFR4, heavy chain FRs including
HFR1, HFR2, HFR3, and HFR4), which are more highly conserved than
the CDRs and form a scaffold to support the highly variable loops.
The constant regions of the heavy and light chains are not involved
in antigen-binding, but exhibit various effector functions.
Antibodies are assigned to classes based on the amino acid
sequences of the constant regions of their heavy chains. The five
major classes or isotypes of antibodies are IgA, IgD, IgE, IgG, and
IgM, which are characterized by the presence of alpha, delta,
epsilon, gamma, and mu heavy chains, respectively. Several of the
major antibody classes are divided into subclasses such as IgG1
(gammal heavy chain), IgG2 (gamma2 heavy chain), IgG3 (gamma3 heavy
chain), IgG4 (gamma4 heavy chain), IgA1 (alpha1 heavy chain), or
IgA2 (alpha2 heavy chain).
[0213] The term "antigen-binding fragment" as used herein refers to
an antibody fragment formed from a portion of an antibody
comprising one or more CDRs, or any other antibody fragment that
binds to an antigen but does not comprise an intact native antibody
structure. Examples of antigen-binding fragment include, without
limitation, a diabody, a Fab, a Fab', a F(ab').sub.2, a Fd, an Fv
fragment, a disulfide stabilized Fv fragment (dsFv), a
(dsFv).sub.2, a bispecific dsFv (dsFv-dsFv'), a disulfide
stabilized diabody (ds diabody), a single-chain antibody molecule
(scFv), an scFv dimer (bivalent diabody), a bispecific scFv dimer,
a single-chain Fv-Fc antibody (scFv-Fc), a camelized single domain
antibody, a nanobody, a domain antibody, and a bivalent domain
antibody. An antigen-binding fragment is capable of binding to the
same antigen to which the parent antibody binds.
[0214] As used herein, a "bispecific" antibody refers to an
artificial antibody which has fragments derived from two different
monoclonal antibodies. A bispecific antibody may bind to
overlapping epitopes or to two different epitopes. The two epitopes
may present on the same antigen, or they may present on two
different antigens. As such, the terms "multi-specific" antibody
refers to an artificial antibody which has fragments derived from
multiple different monoclonal antibodies, and may be capable of
binding to more than one epitope.
[0215] The term "chimeric" as used herein, means an antibody or
antigen-binding fragment, having a portion of heavy and/or light
chain derived from one species, and the rest of the heavy and/or
light chain derived from a different species.
[0216] The term "epitope" as used herein refers to the specific
group of atoms or amino acids on an antigen to which an antibody
binds. Two antibodies may bind the same or a closely related
epitope within an antigen if they exhibit competitive binding for
the antigen. An epitope can be linear or conformational (i.e.
including amino acid residues spaced apart). For example, if an
antibody or antigen-binding fragment blocks binding of a reference
antibody to the antigen by at least 50%, at least 55%, at least
60%, at least 65%, at least 70%, at least 75%, at least 80%, at
least 85%, or at least 90%, or at least 95%, then the antibody or
antigen-binding fragment may be considered to bind the same/closely
related epitope as the reference antibody. The capacity to block,
or compete with, the binding of the antibody or the antigen-binding
fragment of the present disclosure to SARS-CoV-2 typically
indicates that an antibody or the antigen-binding fragment to be
screened binds to an epitope or binding site on SARS-CoV-2 that
structurally overlaps with the binding site on SARS-CoV-2 that is
immunospecifically recognized by the antibody or the
antigen-binding fragment of the present disclosure. Alternatively,
this can indicate that an antibody or an antigen-binding fragment
of the present disclosure to be screened binds to an epitope or
binding site that is sufficiently proximal to the binding site
immunospecifically recognized by the antibody or the
antigen-binding fragment of the present disclosure to sterically or
otherwise inhibit binding of the antibodies or the antigen-binding
fragment of the present disclosure to SARS-CoV-2.
[0217] "Fab" with regard to an antibody refers to that portion of
the antibody consisting of a single light chain (both variable and
constant regions) bound to the variable region and first constant
region of a single heavy chain by a disulfide bond. The heavy chain
fragment of the Fab is known as "Fd".
[0218] "Fab'" refers to a Fab fragment that includes a portion of
the hinge region.
[0219] "F(ab').sub.2" refers to a dimer of Fab'.
[0220] "Fc" with regard to an antibody (e.g. of IgG, IgA, or IgD
isotype) refers to that portion of the antibody consisting of the
second and third constant domains of a first heavy chain bound to
the second and third constant domains of a second heavy chain via
disulfide bonding. Fc with regard to antibody of IgM and IgE
isotype further comprises a fourth constant domain. The Fc portion
of the antibody is responsible for various effector functions such
as antibody-dependent cell-mediated cytotoxicity (ADCC),
Antibody-dependent cellular phagocytosis (ADCP) and complement
dependent cytotoxicity (CDC), but does not function in antigen
binding.
[0221] "Fv" with regard to an antibody refers to the smallest
fragment of the antibody to bear the complete antigen binding site.
An Fv fragment consists of the variable region of a single light
chain bound to the variable region of a single heavy chain.
[0222] "Single-chain Fv antibody" or "scFv" refers to an engineered
antibody consisting of a light chain variable region and a heavy
chain variable region connected to one another directly or via a
peptide linker sequence (Huston J S et al. Proc Natl Acad Sci USA,
85:5879(1988)).
[0223] "ScFab" refers to a fusion polypeptide with a Fd linked to a
light chain via a polypeptide linker, resulting in the formation of
a single chain Fab fragment (scFab).
[0224] "Single-chain Fv-Fc antibody" or "scFv-Fc" refers to an
engineered antibody consisting of a scFv connected to the Fc region
of an antibody.
[0225] "Camelized single domain antibody," "heavy chain antibody,"
or "HCAb" refers to an antibody that contains two V.sub.H domains
and no light chains (Riechmann L. and Muyldermans S., J Immunol
Methods. December 10; 231(1-2):25-38 (1999); Muyldermans S., J
Biotechnol. June; 74(4):277-302 (2001); WO94/04678; WO94/25591;
U.S. Pat. No. 6,005,079). Heavy chain antibodies were originally
derived from Camelidae (camels, dromedaries, and llamas). Although
devoid of light chains, camelized antibodies have an authentic
antigen-binding repertoire (Hamers-Casterman C. et al., Nature.
June 3; 363(6428):446-8 (1993); Nguyen V K. et al. Immunogenetics.
April; 54(1):39-47 (2002); Nguyen V K. et al. Immunology. May;
109(1):93-101 (2003)). The variable domain of a heavy chain
antibody (VHH domain) represents the smallest known antigen-binding
unit generated by adaptive immune responses (Koch-Nolte F. et al.,
FASEB J. November; 21(13):3490-8. Epub 2007 Jun. 15 (2007)).
[0226] A "nanobody" refers to an antibody fragment that consists of
a VHH domain from a heavy chain antibody and two constant domains,
CH2 and CH3.
[0227] A "domain antibody" refers to an antibody fragment
containing only the variable region of a heavy chain or the
variable region of a light chain. In certain instances, two or more
VH domains are covalently joined with a peptide linker to create a
bivalent or multivalent domain antibody. The two VH domains of a
bivalent domain antibody may target the same or different
antigens.
[0228] The term "valent" as used herein refers to the presence of a
specified number of antigen binding sites in a given molecule. The
term "monovalent" refers to an antibody or an antigen-binding
fragment having only one single antigen-binding site; and the term
"multivalent" refers to an antibody or an antigen-binding fragment
having multiple antigen-binding sites. As such, the terms
"bivalent", "tetravalent", and "hexavalent" denote the presence of
two binding sites, four binding sites, and six binding sites,
respectively, in an antigen-binding molecule. In some embodiments,
the antibody or antigen-binding fragment thereof is bivalent.
[0229] "TriFabs" refers to a trivalent, bispecific fusion protein
composed of three units with Fab-functionalities. TriFabs harbor
two regular Fabs fused to an asymmetric Fab-like moiety.
[0230] "Fab-Fab" refers to a fusion protein formed by fusing the Fd
chain of a first Fab arm to the N-terminus of the Fd chain of a
second Fab arm.
[0231] "Fab-Fv" refers to a fusion protein formed by fusing a heavy
chain variable domain to the C-terminus of an Fd chain and a light
chain variable domain to the C-terminus of a light chain. A
"Fab-dsFv" molecule can be formed by introducing an interdomain
disulphide bond between the heavy chain variable domain and the
heavy chain variable domain.
[0232] An "scFv dimer" is a bivalent diabody or bispecific scFv
(BsFv) comprising V.sub.H-V.sub.L (linked by a peptide linker)
dimerized with another V.sub.H-V.sub.L moiety such that V.sub.H's
of one moiety coordinate with the V.sub.L's of the other moiety and
form two binding sites which can target the same antigens (or
epitopes) or different antigens (or epitopes).
[0233] A bispecific "scFv dimer" is a bispecific diabody comprising
V.sub.H1-V.sub.L2 (linked by a peptide linker) associated with
V.sub.L1-V.sub.H2 (also linked by a peptide linker) such that
V.sub.H1 and V.sub.L1 coordinate and V.sub.H2 and V.sub.L2
coordinate and each coordinated pair has a different antigen
specificity.
[0234] A "dsFv" refers to a disulfide-stabilized Fv fragment that
the linkage between the variable region of a single light chain and
the variable region of a single heavy chain is a disulfide bond. In
some embodiments, a "(dsFv).sub.2" or "(dsFv-dsFv')" comprises
three peptide chains: two V.sub.H moieties linked by a peptide
linker (e.g. a long flexible linker) and bound to two V.sub.L
moieties, respectively, via disulfide bridges. In some embodiments,
dsFv-dsFv' is bispecific in which each disulfide paired heavy and
light chain has a different antigen specificity.
[0235] "Bibody" refers to a fusion protein formed by fusing a scFv
to the C-terminus of either the light chain (Fab-L-scFv) or Fd
(Fab-H-scFv).
[0236] "Tribody" refers to a fusion protein formed by fusing a scFv
to both light chain and heavy chain (Fab-(scFv).sub.2).
[0237] "MAb-Fv" or "IgG-Fv" refers to a fusion protein formed by
fusion of heavy chain variable domain (VH domain) to the C-terminus
of one Fc chain and the VL domain either expressed separately or
fused to the C-terminus of the other resulted in a bispecific,
trivalent IgG-Fv (mAb-Fv) fusion protein, with the Fv stabilized by
an interdomain disulphide bond.
[0238] "ScFab-Fc-scFv.sub.2" and "ScFab-Fc-scFv" refer to a fusion
protein formed by fusion of a single-chain Fab with Fc and
disulphide-stabilized Fv domains.
[0239] "Appended IgG" refers to a fusion protein with a Fab arm
fused to an IgG to form the format of bispecific (Fab).sub.2-Fc. It
can form an "IgG-Fab" or a "Fab-IgG", with a Fab fused to the
C-terminus or N-terminus of an IgG molecule with or without a
connector. In certain embodiments, the appended IgG can be further
modified to a format of IgG-Fab.sub.4 (see, Brinkman et al., mAbs,
9(2), pp.182-212 (2017)).
[0240] "DVD-Ig" refers to a dual-variable-domain antibody that is
formed by fusion of an additional VH domain and VL domain of a
second specificity to an IgG heavy chain and light chain. "CODV-Ig"
refers to a related format where the two VH domain and two VL
domains are linked in a way that allows crossover pairing of the
variable VH domain-VL domain, which are arranged either (from N- to
C-terminus) in the order VH domain A-VH domain B and VL domain B-VL
domain A, or in the order VH domain B-VH domain A and VL domain
A-VL domain B.
[0241] A "CrossMab" refers to a technology of pairing of unmodified
light chain with the corresponding unmodified heavy chain and
pairing of the modified light chain with the corresponding modified
heavy chain, thus resulting an antibody with reduced mispairing in
the light chain.
[0242] A "WuxiBody" refers to is a bispecific antibody comprising a
chimeric protein with variable domains of an antibody and the
constant domains of TCR, wherein the subunits (such as alpha and
beta domains) of TCR constant domains are associated by engineered
disulfide bond (see, more details in WO2019057122A1).
[0243] A "BiTE" is a bispecific T-cell engager molecule, comprising
a first scFv with a first antigen specificity in the VL domain-VH
domain orientation linked to a second scFv with a second
specificity in the VH domain-VL domain orientation.
[0244] A "diabody" or "dAb" includes small antibody fragments with
two antigen-binding sites, wherein the fragments comprise a V.sub.H
domain connected to a V.sub.L domain in the same polypeptide chain
(V.sub.H-V.sub.L or VL-VH) (see, e.g. Holliger P. et al., Proc Natl
Acad Sci USA. July 15; 90(14):6444-8 (1993); EP404097; WO93/11161).
By using a linker that is too short to allow pairing between the
two domains on the same chain, the domains are forced to pair with
the complementary domains of another chain, thereby creating two
antigen-binding sites. The antigen-binding sites may target the
same or different antigens (or epitopes).
[0245] A "DART" is a diabody-like entity that has the VH of a first
variable region linked to the VL of a second variable region, and
the VH of the second variable region linked to the VL of the first
variable region.
[0246] A "TandAb" is a bispecific fusion protein with four binding
sites, two of which bind to a first antigen and the other two bind
to a second antigen.
[0247] A "bispecific ds diabody" is a diabody target two different
antigens (or epitopes).
[0248] The term "fully human" when used with reference to an
antibody, refers to an antibody that are either directly derived
from a human or based upon a human sequence. When an antibody is
derived from or based on a human sequence and subsequently
modified, it is still to be considered fully human as used
throughout the specification. In other words, the term "fully
human" when used with reference to an antibody, is intended to
include binding molecules having variable and constant regions
derived from human germline immunoglobulin sequences or based on
variable or constant regions occurring in a human or human
lymphocyte and modified in some form. Thus, the fully human
antibody may include amino acid residues not encoded by human
germline immunoglobulin sequences, comprise substitutions and/or
deletions (e.g., mutations introduced by, for instance, random or
site-specific mutagenesis in vitro or by somatic mutation in vivo).
"Based on" as used herein refers to the situation that a nucleic
acid sequence may be exactly copied from a template, or with minor
mutations, such as by error-prone PCR methods, or synthetically
made matching the template exactly or with minor modifications.
Semi-synthetic molecules based on human sequences are also
considered to be human as used herein.
[0249] Other Terms
[0250] The term "affinity" as used herein refers to the strength of
non-covalent interaction between an immunoglobulin molecule (i.e.
antibody) or fragment thereof and an antigen.
[0251] The term "amino acid" as used herein refers to an organic
compound containing amine (--NH.sub.2) and carboxyl (--COOH)
functional groups, along with a side chain specific to each amino
acid. The names of amino acids are also represented as standard
single letter or three-letter codes in the present disclosure,
which are summarized as follows.
TABLE-US-00001 Three- Single- Name of Amino letter letter Acid Code
Code Alanine Ala A Arginine Arg R Asparagine Asn N Aspartic acid
Asp D Cysteine Cys C Glutamic acid Glu E Glutamine Gln Q Glycine
Gly G Histidine His H Isoleucine Ile I Leucine Leu L Lysine Lys K
Methionine Met M Phenylalanine Phe F Proline Pro P Serine Ser S
Threonine Thr T Tryptophan Trp W Tyrosine Tyr Y Valine Val V
[0252] A "conservative substitution" with reference to amino acid
sequence refers to replacing an amino acid residue with a different
amino acid residue having a side chain with similar physiochemical
properties. For example, conservative substitutions can be made
among amino acid residues with hydrophobic side chains (e.g. Met,
Ala, Val, Leu, and Ile), among residues with neutral hydrophilic
side chains (e.g. Cys, Ser, Thr, Asn and Gln), among residues with
acidic side chains (e.g. Asp, Glu), among amino acids with basic
side chains (e.g. His, Lys, and Arg), or among residues with
aromatic side chains (e.g. Trp, Tyr, and Phe). As known in the art,
conservative substitution usually does not cause significant change
in the protein conformational structure, and therefore could retain
the biological activity of a protein.
[0253] The term "diagnosis", "diagnose" or "diagnosing" refers to
the identification of a pathological state, disease or condition,
such as identification of SARS-CoV-2 infection, or refer to
identification of a subject with SARS-CoV-2 infection who may
benefit from a particular treatment regimen. In some embodiments,
diagnosis contains the identification of presence or amount of
SARS-CoV-2. In some embodiments, diagnosis refers to the
identification of SARS-CoV-2 infection in a subject.
[0254] "Effector functions" as used herein refer to biological
activities attributable to the binding of Fc region of an antibody
to its effectors such as C1 complex and Fc receptor. Exemplary
effector functions include: complement dependent cytotoxicity (CDC)
mediated by interaction of antibodies and C1q on the C1 complex;
antibody-dependent cell-mediated cytotoxicity (ADCC) mediated by
binding of Fc region of an antibody to Fc receptor on an effector
cell; and phagocytosis. Effector functions can be evaluated using
various assays such as Fc receptor binding assay, C1q binding
assay, and cell lysis assay.
[0255] The term "Antibody-dependent cell-mediated cytotoxicity" and
"ADCC" refer to a cell-mediated reaction in which nonspecific
cytotoxic cells that express FcRs (e.g. Natural Killer (NK) cells,
neutrophils, and macrophages) recognize bound antibody on a target
ceil and subsequently cause lysis of the target cell. The primary
cells for mediating ADCC, NK cells, express Fc.gamma.RIII only,
whereas monocytes express Fc.gamma.RI, Fc.gamma.RII and
Fc.gamma.RIII. FcR expression on hematopoietic cells is summarized
in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol
9:457-92 (1991).
[0256] The term "specific binding" or "specifically binds" in
reference to the interaction of a binding molecule, e.g., an
antibody, and its binding partner, e.g., an antigen, means that the
interaction is dependent upon the presence of a particular
structure, e.g., an antigenic determinant or epitope, on the
binding partner. In other words, the antibody preferentially binds
or recognizes the binding partner even when the binding partner is
present in a mixture of other molecules or organisms. The binding
may be mediated by covalent or non-covalent interactions or a
combination of both. Antibodies or fragments thereof that
immunospecifically bind to an antigen may be cross-reactive with
related antigens, carrying the same epitope. Specific binding can
be characterized in binding affinity, for example, represented by
K.sub.d value, i.e., the dissociation constant between the antigen
and antigen-binding molecule. K.sub.d may be determined by using
any conventional method known in the art, including but are not
limited to radioimmunoassays (RIA), enzyme-linked immunosorbent
assays (ELISA), surface plasmon resonance method, microscale
thermophoresis method, HPLC-MS method and flow cytometry (such as
FACS) method. A K.sub.d value of 10.sup.-6 M (e.g.
5.times.10.sup.-7 M, 2.times.10.sup.-7 M, 10.sup.-7 M,
5.times.10.sup.-8 M, 2.times.10.sup.-8 M, 10.sup.-8 M,
5.times.10.sup.-9 M, 4.times.10.sup.-9M, 3.times.10.sup.-9M,
2.times.10.sup.-9 M, or 10.sup.-9 M) can indicate specific binding
between an antibody or antigen binding fragments thereof and
SARS-CoV-2 (e.g. spike protein of SARS-CoV-2, or receptor binding
domain of the spike protein of SARS-CoV-2).
[0257] The ability to "compete for binding to RBD" as used herein
refers to the ability of a SARS-CoV-2 antibody or antigen-binding
fragment thereof to inhibit the binding interaction between RBD of
spike protein of SARS-CoV-2 and its binding partner (e.g. a second
SARS-CoV-2 antibody, or ACE2 receptor) to any detectable degree. In
certain embodiments, an antibody or antigen-binding fragment that
compete for binding to SARS-CoV-2 inhibits the binding interaction
between RBD of spike protein of SARS-CoV-2 and its binding partner
by at least 85%, or at least 90%. In certain embodiments, this
inhibition may be greater than 95%, or greater than 99%. In
general, competitive inhibition is measured by means of an assay,
wherein an antigen composition, i.e., a composition comprising
SARS-CoV-2 or fragments thereof, is admixed with reference binding
molecules, for example, the antibodies or antigen binding fragments
of the present disclosure, or the ACE receptor (e.g. a recombinant
binding moiety thereof), and the antibodies or antigen binding
fragments to be screened. Usually, the antibodies or antigen
binding fragments to be screened are present in excess. Protocols
based upon ELISAs and Western blotting are suitable for use in such
simple competition studies.
[0258] In certain embodiments, an antibody or antigen-binding
fragment exhibits at least 30% competition at 1 .mu.M, with 2 .mu.M
angiotensin converting enzyme 2 (ACE2) receptor for binding to the
RBD of spike protein of SARS-CoV-2 immobilized at a resonance units
(RU) of 250, as measured by SPR.
[0259] The term "homologous" as used herein refers to nucleic acid
sequences (or its complementary strand) or amino acid sequences
that have sequence identity of at least 60% (e.g. at least 65%,
70%, 75%, 80%, 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) to another sequences when optimally aligned.
[0260] The phrase "host cell" as used herein refers to a cell into
which an exogenous polynucleotide and/or a vector can be or has
been introduced.
[0261] The term "isolated" means one substance has been altered by
the hand of man from the natural state. If an "isolated"
composition or substance occurs in nature, it has been changed or
removed from its original environment, or both. For example, a
polynucleotide or a polypeptide naturally present in a living
animal is not "isolated," but the same polynucleotide or
polypeptide is "isolated" if it has been sufficiently separated
from the coexisting materials of its natural state so as to exist
in a substantially pure state. An "isolated nucleic acid sequence"
refers to the sequence of an isolated nucleic acid molecule. In
certain embodiments, an "isolated antibody or an antigen-binding
fragment thereof" refers to the antibody or antigen-binding
fragments thereof having a purity of at least 60%, 70%, 75%, 80%,
81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%,
94%, 95%, 96%, 97%, 98%, 99% as determined by electrophoretic
methods (such as SDS-PAGE, isoelectric focusing, capillary
electrophoresis), or chromatographic methods (such as ion exchange
chromatography or reverse phase HPLC). In some embodiment, an
isolated antibody or antigen binding fragment is a recombinant
protein or antigen binding fragment.
[0262] The term "modified antibody", "modified antibodies", or a
grammatic variation as used herein refers to an antibody that has
been modified, bioengineered, or combined with one or more
modification elements so it is not a naturally occurring
antibody.
[0263] The term "kit" as used herein refers to a packaged
combination of reagents in predetermined amounts with instructions
for performing a therapeutics, or a diagnostic or detection
assay.
[0264] The term "neutralizing" as used herein in relation to the
antibody or the antigen binding fragment of the present disclosure
refers to antibody or the antigen binding fragment that inhibit
SARS-CoV-2 virus from infecting a target cell for replication,
regardless of the mechanism by which neutralization is achieved.
Thus, neutralization can, for example, be achieved by inhibiting
the attachment or adhesion of SARS-CoV-2 virus or a pseudo
SARS-CoV-2 virus bearing the spike protein to the cell surface, or
by inhibition of the fusion of viral and cellular membranes
following attachment of the virus to the target cell, and the like.
Exemplary assays for determining neutralizing activity are
described in the Examples provided herein.
[0265] In some embodiments, the neutralizing activity of an
antibody can be represented as half-maximal inhibitory
concentrations (IC.sub.50) of the antibody against the binding to
ACE2.
[0266] The term "nucleic acid" or "polynucleotide" as used herein
refers to deoxyribonucleic acids (DNA) or ribonucleic acids (RNA)
and polymers thereof in either single- or double-stranded form.
Unless otherwise indicated, a particular polynucleotide sequence
also implicitly encompasses conservatively modified variants
thereof (e.g. degenerate codon substitutions), alleles, orthologs,
SNPs, and complementary sequences as well as the sequence
explicitly indicated. Specifically, degenerate codon substitutions
may be achieved by generating sequences in which the third position
of one or more selected (or all) codons is substituted with
mixed-base and/or deoxyinosine residues (see Batzer et al., Nucleic
Acid Res. 19:5081 (1991); Ohtsuka et al., J. Biol. Chem.
260:2605-2608 (1985); and Rossolini et al., Mol. Cell. Probes
8:91-98 (1994)).
[0267] "Percent (%) sequence identity" with respect to amino acid
sequence (or nucleic acid sequence) is defined as the percentage of
amino acid (or nucleic acid) residues in a candidate sequence that
are identical to the amino acid (or nucleic acid) residues in a
reference sequence, after aligning the sequences and, if necessary,
introducing gaps, to achieve the maximum number of identical amino
acids (or nucleic acids). Conservative substitution of the amino
acid residues may or may not be considered as identical residues.
Alignment for purposes of determining percent amino acid (or
nucleic acid) sequence identity can be achieved, for example, using
publicly available tools such as BLASTN, BLASTp (available on the
website of U.S. National Center for Biotechnology Information
(NCBI), see also, Altschul S. F. et al., J. Mol. Biol., 215:403-410
(1990); Stephen F. et al., Nucleic Acids Res., 25:3389-3402
(1997)), ClustalW2 (available on the website of European
Bioinformatics Institute, see also, Higgins D. G. et al., Methods
in Enzymology, 266:383-402 (1996); Larkin M. A. et al.,
Bioinformatics (Oxford, England), 23(21): 2947-8 (2007)), and ALIGN
or Megalign (DNASTAR) software. A person skilled in the art may use
the default parameters provided by the tool, or may customize the
parameters as appropriate for the alignment, such as for example,
by selecting a suitable algorithm.
[0268] The term "polypeptide" or "protein" means a string of at
least two amino acids linked to one another by peptide bonds.
Polypeptides and proteins may include moieties in addition to amino
acids (e.g., may be glycosylated) and/or may be otherwise processed
or modified. Those of ordinary skill in the art will appreciate
that a "polypeptide" or "protein" can be a complete polypeptide
chain as produced by a cell (with or without a signal sequence), or
can be a functional portion thereof. Those of ordinary skill will
further appreciate that a polypeptide or protein can sometimes
include more than one polypeptide chain, for example linked by one
or more disulfide bonds or associated by other means. The term also
includes amino acid polymers in which one or more amino acids are
chemical analogs of a corresponding naturally-occurring amino acid
and polymers.
[0269] The term "pharmaceutically acceptable" indicates that the
designated carrier, vehicle, diluent, excipient(s), and/or salt is
generally chemically and/or physically compatible with the other
ingredients comprising the formulation, and physiologically
compatible with the recipient thereof.
[0270] The term "recombinant" when used with reference to a
polypeptide (e.g., antibody, antigen) or a polynucleotide, refers
to a polypeptide or polynucleotide that is produced by a
recombinant method. A "recombinant polypeptide" includes any
polypeptide expressed from a recombinant polynucleotide. A
"recombinant polynucleotide" includes any polynucleotide which has
been modified by the introduction of at least one exogenous (i.e.,
foreign, and typically heterologous) nucleotide or the alteration
of at least one native nucleotide component of the polynucleotide,
and need not include all of the coding sequence or the regulatory
elements naturally associated with the coding sequence. A
"recombinant vector" refers to a non-naturally occurring vector,
including, e.g., a vector comprising a recombinant polynucleotide
sequence.
[0271] As used herein, the term "sample" refers to a biological
specimen that is obtained or derived from a subject of interest.
The sample contains a cellular and/or other molecular entity that
is to be characterized and/or identified, for example based on
physical, biochemical, chemical and/or physiological
characteristics.
[0272] The term "subject" includes human and non-human animals.
Non-human animals include all vertebrates, e.g., mammals and
non-mammals, such as non-human primates, mice, rats, cats, rabbits,
sheep, dogs, cows, chickens, amphibians, and reptiles. Except when
noted, the terms "patient" or "subject" are used herein
interchangeably.
[0273] The term "treating" or "treatment" of a disease, disorder or
condition as used herein includes alleviating a disease, disorder
or condition, slowing the rate of development of a disease,
disorder or condition, reducing or ending symptoms associated with
a disease, disorder or condition, generating a complete or partial
regression of a disease, disorder or condition, curing a disease,
disorder or condition, or some combination thereof.
[0274] The term "prevent" or "preventing" of a disease, disorder or
condition as used herein includes slowing the onset of a disease,
disorder or condition, reducing the risk of developing a disease,
disorder or condition, preventing or delaying the development of
symptoms associated with a disease, disorder or condition, reducing
the severity of a subsequent contraction or development of a
disease, disorder or condition, ameliorating a related symptom, and
inducing immunity to protect against a disease, disorder or
condition.
[0275] The term "SARS-CoV-2 virus antigen" as used herein refers to
a SARS-CoV-2 virus particle or an antigen found in a SARS-CoV-2
virus particle (e.g. a protein or protein fragments of envelop
protein or spike protein (includes, extracellular domain of the
spike protein, or RBD of the spike protein) and the like). Spike
protein is composed of S1 protein (which contains RBD) and S2
protein, which are initially in one protein molecule until cleaved
by protease into S1 and S2.
[0276] The term "vector" as used herein refers to a vehicle into
which a genetic element may be operably inserted so as to bring
about the expression of that genetic element, such as to produce
the protein, RNA or DNA encoded by the genetic element, or to
replicate the genetic element. A vector may be used to transform,
transduce, or transfect a host cell so as to bring about expression
of the genetic element it carries within the host cell. Examples of
vectors include plasmids, phagemids, cosmids, artificial
chromosomes such as yeast artificial chromosome (YAC), bacterial
artificial chromosome (BAC), or P1-derived artificial chromosome
(PAC), bacteriophages such as lambda phage or M13 phage, and animal
viruses. A vector may contain a variety of elements for controlling
expression, including promoter sequences, transcription initiation
sequences, enhancer sequences, selectable elements, and reporter
genes. In addition, the vector may contain an origin of
replication. A vector may also include materials to aid in its
entry into the cell, including but not limited to a viral particle,
a liposome, or a protein coating. A vector can be an expression
vector or a cloning vector. The present disclosure provides vectors
(e.g. expression vectors) containing the nucleic acid sequence
provided herein encoding the antibody or an antigen-binding
fragment thereof, at least one promoter (e.g. SV40, CMV,
EF-1.alpha.) operably linked to the nucleic acid sequence, and at
least one selection marker.
[0277] Anti-SARS-CoV-2 Antibodies
[0278] The present disclosure in one aspect provides
anti-SARS-CoV-2 antibodies and antigen-binding fragments
thereof.
[0279] In some embodiments, the disclosure is directed to a
modified antibody or an antigen-binding fragment thereof comprising
at least an antigen-binding domain having an antigen-binding
affinity and a covalently linked modified human IgG constant
domain, wherein the antigen-binding affinity comprises SARS-CoV-2
binding affinity, the antigen-binding affinity comprises at least
50% less or non-detectable binding affinity to SARS-CoV or MERS-CoV
compared to the SARS-CoV-2 binding affinity, and wherein the
modified human IgG constant domain comprises a substitution with
tyrosine at amino acid residue 252, a substitution with threonine
at amino acid residue 254, and a substitution with glutamic acid at
amino acid residue 256, numbered according to the EU index as in
Kabat, the modified antibody has an increased affinity for FcRn
compared to the affinity to FcRn of an antibody having a wild type
human IgG constant domain.
[0280] The modified human IgG constant domain comprises a
substitution with tyrosine at amino acid residue 252, a
substitution with threonine at amino acid residue 254, and a
substitution with glutamic acid at amino acid residue 256, numbered
according to the EU index as in Kabat, can be referred to YTE
domain or YTE domain Fc.
[0281] In some cases, the antigen-binding affinity can
comprise:
[0282] a) binding affinity to spike protein of SARS-CoV-2 with at
least 50% less binding to spike protein of SARS-CoV or spike
protein of MERS-CoV;
[0283] b) binding affinity to receptor-binding domain (RBD) of the
spike protein of SARS-CoV-2 comprising the amino acid sequence of
SEQ ID NO: 128;
[0284] c) binding affinity to RBD of said spike protein of SARS-CoV
comprising the amino acid sequence of SEQ ID NO: 124 at a level
that is non-detectable or that is no more than 50% of said binding
affinity to said RBD of said spike protein of SARS-CoV-2;
[0285] d) binding affinity to RBD of said spike protein of MERS-CoV
comprising the amino acid sequence of SEQ ID NO: 126 at a level
that is non-detectable or that is no more than 50% of said binding
affinity to said RBD of the spike protein of SARS-CoV-2;
[0286] e) binding affinity to said RBD of said spike protein of
SARS-CoV-2 at a Kd value of no more than 1.times.10.sup.-7M as
measured by Surface Plasmon Resonance (SPR);
[0287] f) binding affinity to said RBD of said spike protein of
SARS-CoV or the RBD of spike protein of MERS-CoV at a Kd value of
at least 1.times.10.sup.-6M as measured by SPR;
[0288] g) exhibiting at least 30% competition at 104, with 2 .mu.M
angiotensin converting enzyme 2 (ACE2) receptor, for binding to
said RBD of said spike protein of SARS-CoV-2 immobilized at a
resonance unit (RU) of 250, as measured by SPR;
[0289] h) binding affinity to said RBD of said spike protein of
SARS-CoV-2 at a neutralizing activity at an IC50 value of no more
than 100 .mu.g/ml (for example, no more than 50 .mu.g/ml, no more
than 40 .mu.g/ml, no more than 30 .mu.g/ml, no more than 25
.mu.g/ml, no more than 20 .mu.g/ml, no more than 15 .mu.g/ml, no
more than 10 .mu.g/ml, no more than 8 .mu.g/ml, no more than 6
.mu.g/ml, no more than 4 .mu.g/ml, no more than 2 .mu.g/ml, or no
more than 1 .mu.g/ml), as measured by pseudovirus, live virus
microneutralization, inactivated virus neutralization assay, or a
combination thereof;
[0290] i) capable of binding to the RBD of spike protein of
SARS-CoV-2 at an neutralizing activity at an IC.sub.50 value of no
more than 1 .mu.g/ml (for example, no more than 50 ng/ml, no more
than 40 ng/ml, no more than 30 ng/ml, no more than 25 ng/ml, no
more than 20 ng/ml, no more than 15 ng/ml, no more than 10 ng/ml,
no more than 8 ng/ml, no more than 6 ng/ml, no more than 4 ng/ml,
no more than 2 ng/ml, or no more than 1 ng/ml), as measured by live
virus neutralization assay using focus reduction neutralization
test (FRNT) method or
[0291] a combination thereof.
[0292] In some cases, the antigen-binding affinity can be selected
from the group consisting of:
[0293] a) binding affinity to spike protein of SARS-CoV-2 with at
least 50% less binding to spike protein of SARS-CoV or spike
protein of MERS-CoV;
[0294] b) binding affinity to receptor-binding domain (RBD) of the
spike protein of SARS-CoV-2 comprising the amino acid sequence of
SEQ ID NO: 128;
[0295] c) binding affinity to RBD of said spike protein of SARS-CoV
comprising the amino acid sequence of SEQ ID NO: 124 at a level
that is non-detectable or that is no more than 50% of said binding
affinity to said RBD of said spike protein of SARS-CoV-2;
[0296] d) binding affinity to RBD of said spike protein of MERS-CoV
comprising the amino acid sequence of SEQ ID NO: 126 at a level
that is non-detectable or that is no more than 50% of said binding
affinity to said RBD of the spike protein of SARS-CoV-2;
[0297] e) binding affinity to said RBD of said spike protein of
SARS-CoV-2 at a Kd value of no more than 1.times.10-7M as measured
by Surface Plasmon Resonance (SPR);
[0298] f) binding affinity to said RBD of said spike protein of
SARS-CoV or the RBD of spike protein of MERS-CoV at a Kd value of
at least 1.times.10-6M as measured by SPR;
[0299] g) exhibiting at least 30% competition at 1 .mu.M, with 2
.mu.M angiotensin converting enzyme 2 (ACE2) receptor, for binding
to said RBD of said spike protein of SARS-CoV-2 immobilized at a
resonance unit (RU) of 250, as measured by SPR;
[0300] h) binding affinity to said RBD of said spike protein of
SARS-CoV-2 at a neutralizing activity at an IC50 value of no more
than 100 .mu.g/ml, as measured by pseudovirus, live virus
microneutralization, inactivated virus neutralization assay, or a
combination thereof;
[0301] i) capable of binding to the RBD of spike protein of
SARS-CoV-2 at an neutralizing activity at an IC.sub.50 value of no
more than 1 .mu.g/ml (for example, no more than 50 ng/ml, no more
than 40 ng/ml, no more than 30 ng/ml, no more than 25 ng/ml, no
more than 20 ng/ml, no more than 15 ng/ml, no more than 10 ng/ml,
no more than 8 ng/ml, no more than 6 ng/ml, no more than 4 ng/ml,
no more than 2 ng/ml, or no more than 1 ng/ml), as measured by live
virus neutralization assay using focus reduction neutralization
test (FRNT) method;
[0302] and a combination thereof.
[0303] In some embodiments, the anti-SARS-CoV-2 antibodies and
antigen-binding fragments provided herein are capable of
specifically binding to SARS-CoV-2. In certain embodiments, the
antibodies and the antigen-binding fragments thereof provided
herein specifically bind to SARS-CoV-2 at an Kd value of no more
than 10.sup.-7 M as measured by SPR.
[0304] In certain embodiments, the antibodies and the
antigen-binding fragments thereof provided herein are capable of
binding to the RBD of spike protein of SARS-CoV-2 at a Kd value of
no more than 1.times.10.sup.-7M (e.g. no more than
5.times.10.sup.-7 M, no more than 2.times.10.sup.-7 M, no more than
10.sup.-7 M, no more than 5.times.10.sup.-8 M, no more than
2.times.10.sup.-8 M, no more than 10.sup.-8 M, no more than
5.times.10.sup.-9 M, no more than 4.times.10.sup.-9M, no more than
3.times.10.sup.-9M, no more than 2.times.10.sup.-9 M, or no more
than 10.sup.-9 M) as measured by SPR.
[0305] In certain embodiments, the antibodies and the
antigen-binding fragments thereof provided herein bind to the RBD
of spike protein of SARS-CoV or the RBD of spike protein of
MERS-CoV at a significantly lower affinity or degree. In certain
embodiments, the antibodies and the antigen-binding fragments
thereof provided herein exhibit binding to the RBD of spike protein
of SARS-CoV or the RBD of spike protein of MERS-CoV at a Kd value
of at least 1.times.10.sup.-6M (e.g. at least 2.times.10.sup.-6 M,
at least 5.times.10.sup.-6 M, at least 10.sup.-5 M) as measured by
SPR.
[0306] In certain embodiments, the antibodies and the
antigen-binding fragments thereof provided herein do not detectably
bind to SARS-CoV or MERS-CoV. In certain embodiments, the
antibodies and the antigen-binding fragments thereof provided
herein exhibits at least 50% (e.g., at least 55%, at least 60%, at
least 65%, at least 70%, at least 75%, at least 80%, at least 85%,
at least 90%, at least 95%, at least 96%, at least 97%, at least
98%, at least 99%) less binding or non-detectable binding to
SARS-CoV or MERS-CoV, than the binding to SARS-CoV-2 under
equivalent assay conditions. In certain embodiments, the antibodies
and the antigen-binding fragments thereof provided herein are
capable of specifically binding to spike protein of SARS-CoV-2 and
exhibiting at least 50% (e.g., at least 55%, at least 60%, at least
65%, at least 70%, at least 75%, at least 80%, at least 85%, at
least 90%, at least 95%, at least 96%, at least 97%, at least 98%,
at least 99%) less binding to spike protein of SARS-CoV or spike
protein of MERS-CoV, than the binding to spike protein of
SARS-CoV-2 under equivalent assay conditions. In certain
embodiments, the full length of spike protein of SARS-CoV-2 can
comprise an amino acid sequence of SEQ ID NO: 134, optionally
encoded by a polynucleotide sequence of SEQ ID NO: 135. In certain
embodiments, the antibodies and the antigen-binding fragments
thereof provided herein are capable of specifically binding to
receptor-binding domain (RBD) of the spike protein of SARS-CoV-2
comprising the amino acid sequence of SEQ ID NO: 128. In certain
embodiments, the antibodies and the antigen-binding fragments
thereof provided herein exhibit binding to RBD of spike protein of
SARS-CoV comprising the amino acid sequence of SEQ ID NO: 124 at a
level that is non-detectable or that is no more than 50% (e.g., no
more than 45%, 40%, 35%, 30%, 25%, 20%, 15%, 10%, 5%, 2%, 1%) of
the binding to the RBD of spike protein of SARS-CoV-2 under
equivalent assay conditions. In certain embodiments, the antibodies
and the antigen-binding fragments thereof provided herein exhibit
binding to RBD of spike protein of MERS-CoV comprising the amino
acid sequence of SEQ ID NO: 126 at a level that is non-detectable
or that is no more than 50% (e.g., no more than 45%, 40%, 35%, 30%,
25%, 20%, 15%, 10%, 5%, 2%, 1%) of the binding to RBD of the spike
protein of SARS-CoV-2 under equivalent assay conditions.
[0307] In certain embodiments, the antibodies and the
antigen-binding fragments thereof provided herein are capable of
exhibiting at least 30% competition at 1 .mu.M, with 2 .mu.M ACE2
receptor for binding to the RBD of spike protein of SARS-CoV-2
immobilized at a (RU of 250, as measured by SPR. For example,
SARS-CoV-2 RBD can be immobilized to a CM5 sensor chip via amine
group for a final RU around 250. 1 .mu.M of the antibodies or the
antigen-binding fragments thereof provided herein can be injected
onto the chip until binding steady-state is reached. 2 .mu.M of
human ACE2 or human ACE2 peptidase domain can be injected for 60
seconds. Blocking efficacy can be determined by comparison of
response units with and without the antibody incubation.
Instruments and kits for SPR are commercially available as, for
example, Biacore T200, GE Healthcare.
[0308] In certain embodiments, the antibodies and the
antigen-binding fragments thereof provided herein are capable of
binding to the RBD of spike protein of SARS-CoV-2 at an
neutralizing activity at an IC.sub.50 value of no more than 100
.mu.g/ml (e.g., no more than 90 .mu.g/ml, 80 .mu.g/ml, 70 .mu.g/ml,
60 .mu.g/ml, 50 .mu.g/ml, 40 .mu.g/ml, 30 .mu.g/ml, 20 .mu.g/ml, 10
.mu.g/ml, 5 .mu.g/ml, 2 .mu.g/ml, 1 .mu.g/ml, 0.5 .mu.g/ml, 0.2
.mu.g/ml, 0.1 .mu.g/ml, 0.05 .mu.g/ml, 0.03 .mu.g/ml), as measured
by pseudovirus neutralization assay. Pseudovirus neutralization
assay are known in the art, and in general involves generating a
pseudovirus expressing a reporter gene and a viral protein of
interest (such as the full length spike protein of SARS-CoV-2 of
SEQ ID NO: 134). The antibodies and the antigen-binding fragments
thereof provided herein can be incubated with the pseudovirus, and
the titer of the pseudovirus can be determined via the report gene.
IC.sub.50 is the concentration of the antibodies or the
antigen-binding fragment thereof can inhibit 50% of the pseudovirus
titer in the assay.
[0309] Illustrative Anti-SARS-CoV-2 Antibodies
[0310] In certain embodiments, the present disclosure provides
SARS-CoV-2 antibodies and antigen-binding fragments thereof
comprising one or more (e.g. 1, 2, 3, 4, 5, or 6) CDRs comprising
the sequences selected from the group consisting of SEQ ID NO: 1-6,
11-16, 21-26, 31-36, 41-46, 51-56, 65-70, 75-80, 85-90, 95-100,
105-110, 136-141, 146-151, 156-161, 166-171, 176-181, 186-191,
196-201, 206-211, 216-221, 226-231, 236-241, 246-251, 256-261,
266-271, 276-281, 286-291, 296-301, 306-311, 316-321, 326-331,
336-341, 346-351, 356-361, 366-371, 376-381, 386-391, 396-401,
406-411, 416-421, and 426-431.
[0311] Antibody "P2A-1A8" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 7, and a light chain variable region
having the sequence of SEQ ID NO: 8.
[0312] Antibody "P2A-1A9" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 17, and a light chain variable region
having the sequence of SEQ ID NO: 18.
[0313] Antibody "P2A-1A10" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 27, and a light chain variable region
having the sequence of SEQ ID NO: 28.
[0314] Antibody "P2A-1B3" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 37, and a light chain variable region
having the sequence of SEQ ID NO: 38.
[0315] Antibody "P2B-2F6" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 47, and a light chain variable region
having the sequence of SEQ ID NO: 48.
[0316] Antibody "P2B-2G4" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 57, and a light chain variable region
having the sequence of SEQ ID NO: 58.
[0317] Antibody "P2B-2G11" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 61, and a light chain variable region
having the sequence of SEQ ID NO: 62.
[0318] Antibody "P2C-1A3" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 71, and a light chain variable region
having the sequence of SEQ ID NO: 72.
[0319] Antibody "P2C-1C8" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 81, and a light chain variable region
having the sequence of SEQ ID NO: 82.
[0320] Antibody "P2C-1C10" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 91, and a light chain variable region
having the sequence of SEQ ID NO: 92.
[0321] Antibody "P2C-1D5" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 101, and a light chain variable region
having the sequence of SEQ ID NO: 102.
[0322] Antibody "P2C-1F11" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 111, and a light chain variable region
having the sequence of SEQ ID NO: 112.
[0323] Antibody "P2B-1G5" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 142, and a light chain variable region
having the sequence of SEQ ID NO: 143.
[0324] Antibody "P2B-1A1" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 152, and a light chain variable region
having the sequence of SEQ ID NO: 153.
[0325] Antibody "P2C-1D7" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 162, and a light chain variable region
having the sequence of SEQ ID NO: 163.
[0326] Antibody "P2B-1A10" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 172, and a light chain variable region
having the sequence of SEQ ID NO: 173.
[0327] Antibody "P2B-1D9" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 182, and a light chain variable region
having the sequence of SEQ ID NO: 183.
[0328] Antibody "P2B-1E4" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 192, and a light chain variable region
having the sequence of SEQ ID NO: 193.
[0329] Antibody "P2B-1G1" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 202, and a light chain variable region
having the sequence of SEQ ID NO: 203.
[0330] Antibody "P4A-2D9" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 212, and a light chain variable region
having the sequence of SEQ ID NO: 213.
[0331] Antibody "P5A-2G7" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 222, and a light chain variable region
having the sequence of SEQ ID NO: 223.
[0332] Antibody "P5A-3C8" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 232, and a light chain variable region
having the sequence of SEQ ID NO: 233.
[0333] Antibody "P5A-1D2" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 242, and a light chain variable region
having the sequence of SEQ ID NO: 243.
[0334] Antibody "P5A-2F11" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 252, and a light chain variable region
having the sequence of SEQ ID NO: 253.
[0335] Antibody "P5A-2E1" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 262, and a light chain variable region
having the sequence of SEQ ID NO: 263.
[0336] Antibody "P5A-1C8" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 272, and a light chain variable region
having the sequence of SEQ ID NO: 273.
[0337] Antibody "P1A-1C10" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 282, and a light chain variable region
having the sequence of SEQ ID NO: 283.
[0338] Antibody "P4A-1H6" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 292, and a light chain variable region
having the sequence of SEQ ID NO: 293.
[0339] Antibody "P4B-1F4" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 302, and a light chain variable region
having the sequence of SEQ ID NO: 303.
[0340] Antibody "P5A-1B6" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 312, and a light chain variable region
having the sequence of SEQ ID NO: 313.
[0341] Antibody "P5A-1B8" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 322, and a light chain variable region
having the sequence of SEQ ID NO: 323.
[0342] Antibody "P5A-1B9" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 332, and a light chain variable region
having the sequence of SEQ ID NO: 333.
[0343] Antibody "P5A-1D1" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 342, and a light chain variable region
having the sequence of SEQ ID NO: 343.
[0344] Antibody "P5A-1D10" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 352, and a light chain variable region
having the sequence of SEQ ID NO: 353.
[0345] Antibody "P5A-2D11" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 362, and a light chain variable region
having the sequence of SEQ ID NO: 363.
[0346] Antibody "P5A-2G9" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 372, and a light chain variable region
having the sequence of SEQ ID NO: 373.
[0347] Antibody "P5A-2H3" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 382, and a light chain variable region
having the sequence of SEQ ID NO: 383.
[0348] Antibody "P5A-3A1" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 392, and a light chain variable region
having the sequence of SEQ ID NO: 393.
[0349] Antibody "P5A-3A6" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 402, and a light chain variable region
having the sequence of SEQ ID NO: 403.
[0350] Antibody "P5A-3B4" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 412, and a light chain variable region
having the sequence of SEQ ID NO: 413.
[0351] Antibody "P5A-3C12" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 422, and a light chain variable region
having the sequence of SEQ ID NO: 423.
[0352] Antibody "P22A-1D1" as used herein refers to a monoclonal
fully human antibody having a heavy chain variable region having
the sequence of SEQ ID NO: 432, and a light chain variable region
having the sequence of SEQ ID NO: 433.
[0353] Table 1 below shows the CDR amino acid sequences of
antibodies P2A-1A8, P2A-1A9, P2B-2G11, P2A-1A10, P2A-1B3, P2B-2F6,
P2B-2G4, P2C-1A3, P2C-1C8, P2C-1C10, P2C- 1D5, P2C-1F11, P2B-1G5,
P2B-1A1, P2C-1D7, P2B-1A10, P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9,
P5A-2G7, P5A-3C8, P5A-1D2, P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10,
P4A- 1H6, P4B-1F4, P5A-1B6, P5A-1B8, P5A-1B9, P5A-1D1, P5A-1D10,
P5A-2D11, P5A-2G9, P5A-2H3, P5A-3A1, P5A-3A6, P5A-3B4, P5A-3C12,
and P22A-1D1.
TABLE-US-00002 TABLE 1 CDR amino acid sequences of 42 antibodies
CDR1 CDR2 CDR3 P2A-1A8 HCDR SEQ ID NO: 1 SEQ ID NO: 2 SEQ ID NO: 3
GFAFDDYA STWNSGTI AKLGGYSDYDYPR PGDHYYGLDV LCDR SEQ ID NO: 4 SEQ ID
NO: 5 SEQ ID NO: 6 SSDVGSYNL DVN RSYTDSNTYV P2A-1A9 HCDR SEQ ID NO:
11 SEQ ID NO: 12 SEQ ID NO: 13 GFTFDDYA ISWNGGII AKVAGRGDYDYY YGMDV
LCDR SEQ ID NO: 14 SEQ ID NO: 15 SEQ ID NO: 16 SSNIGAGYD GNN
QSYDSSLSGSV P2A-1A10 HCDR SEQ ID NO: 21 SEQ ID NO: 22 SEQ ID NO: 23
GYTFTGYY INPNSGGT ARVPYCSSTSCHRD WYFDL LCDR SEQ ID NO: 24 SEQ ID
NO: 25 SEQ ID NO: 26 QSLLDSDDGNTY TLS MQRIEFPLT P2A-1B3 HCDR SEQ ID
NO: 31 SEQ ID NO: 32 SEQ ID NO: 33 GFSFNRYS ISASGNTI ARPAMVREGTYN
WFDP LCDR SEQ ID NO: 34 SEQ ID NO: 35 SEQ ID NO: 36 QSVSNDY YAS
QQYGDSPPIT P2B-2F6 HCDR SEQ ID NO: 41 SEQ ID NO: 42 SEQ ID NO: 43
GYSISSGYY IYHSGST ARAVVGIVVVPAA GRRAFDI LCDR SEQ ID NO: 44 SEQ ID
NO: 45 SEQ ID NO: 46 SSDVGGYNY EVS SSYAGSNNLV P2B-2G4 HCDR SEQ ID
NO: 51 SEQ ID NO: 52 SEQ ID NO: 53 GFTFSSYG IWYDGSNK ARGAAMVWLDY
LCDR SEQ ID NO: 54 SEQ ID NO: 55 SEQ ID NO: 56 SSDVGGYNY DVS
CSYAGSYTFVV P2B-2G11 HCDR SEQ ID NO: 11 SEQ ID NO: 12 SEQ ID NO: 13
GFTFDDYA ISWNGGII AKVAGRGDYDYY YGMDV LCDR SEQ ID NO: 14 SEQ ID NO:
15 SEQ ID NO: 16 SSNIGAGYD GNN QSYDSSLSGSV P2C-1A3 HCDR SEQ ID NO:
65 SEQ ID NO: 66 SEQ ID NO: 67 GFTFSDYY ISSSGSTI ARDFSHQQLVPS LCDR
SEQ ID NO: 68 SEQ ID NO: 69 SEQ ID NO: 70 QGISSY AAS QQLNSYPLT
P2C-1C8 HCDR SEQ ID NO: 75 SEQ ID NO: 76 SEQ ID NO: 77 GFTFRSYG
IWYDGSNK ARDIEIVVVNIDY LCDR SEQ ID NO: 78 SEQ ID NO: 79 SEQ ID NO:
80 QSLVYSDGNTY KVS MQGTHWPYT P2C-1C10 HCDR SEQ ID NO: 85 SEQ ID NO:
86 SEQ ID NO: 87 GGTFSSYA IIPIFGTA ARVVTGYYFDY LCDR SEQ ID NO: 88
SEQ ID NO: 89 SEQ ID NO: 90 QSVSSY DAS QQRSNWPS P2C-1D5 HCDR SEQ ID
NO: 95 SEQ ID NO: 96 SEQ ID NO: 97 GFTFSSFA ISGSGGST AKDPDGSGSWYFD
Y LCDR SEQ ID NO: 98 SEQ ID NO: 99 SEQ ID NO: 100 NIGSKS YDS
QVWDSSSDHEIV P2C-1F11 HCDR SEQ ID NO: 105 SEQ ID NO: 106 SEQ ID NO:
107 GITVSSNY IYSGGST ARDLVVYGMDV LCDR SEQ ID NO: 108 SEQ ID NO: 109
SEQ ID NO: 110 QSVSSSY GAS QQYGSSPT P2B-1G5 HCDR SEQ ID NO: 136 SEQ
ID NO: 137 SEQ ID NO: 138 GYTFTTYV INTNTGNP SCEITTLGGMDV LCDR SEQ
ID NO: 139 SEQ ID NO: 140 SEQ ID NO: 141 NIGSKS YDS QVWDSISDHRV
P2B-1A1 HCDR SEQ ID NO: 146 SEQ ID NO: 147 SEQ ID NO: 148 GGSISSYY
IYYSGST ARLERDWPLDAFDI LCDR SEQ ID NO: 149 SEQ ID NO: 150 SEQ ID
NO: 151 SSDVGGYNY DVS SSYTSNNTFA P2C-1D7 HCDR SEQ ID NO: 156 SEQ ID
NO: 157 SEQ ID NO: 158 GFTVSSNY IYSGGST ARELYEVGATDY LCDR SEQ ID
NO: 159 SEQ ID NO: 160 SEQ ID NO: 161 QSLVYSDGNTY KVS MQRYTLAGV
P2B-1A10 HCDR SEQ ID NO: 166 SEQ ID NO: 167 SEQ ID NO: 168 GFTVSSNY
IYSGGST AREGPKSITGTAFDI LCDR SEQ ID NO: 169 SEQ ID NO: 170 SEQ ID
NO: 171 QDISNY DAS QQYDNLPMYT P2B-1D9 HCDR SEQ ID NO: 176 SEQ ID
NO: 177 SEQ ID NO: 178 GFSLSTSGVG IYWDDDK AHTRILYYGSGSYY DY LCDR
SEQ ID NO: 179 SEQ ID NO: 180 SEQ ID NO: 181 SSNIGSNY SNN
AAWDDSLSGVV P2B-1E4 HCDR SEQ ID NO: 186 SEQ ID NO: 187 SEQ ID NO:
188 GFSLSTSGVG IYWDDDK AHQIVATIIDY LCDR SEQ ID NO: 189 SEQ ID NO:
190 SEQ ID NO: 191 SSDVGGYNY DVS SSYTSSSVV P2B-1G1 HCDR SEQ ID NO:
196 SEQ ID NO: 197 SEQ ID NO: 198 GFTVSSNY IYSGGST ARDYGDYWFDP LCDR
SEQ ID NO: 199 SEQ ID NO: 200 SEQ ID NO: 201 QSVSSSY GAS QQYGSSPRT
P4A-2D9 HCDR SEQ ID NO: 206 SEQ ID NO: 207 SEQ ID NO: 208 GFTFSSYG
ISDDGSNQ AKRGGYCSTTSCL VRWVYFDY LCDR SEQ ID NO: 209 SEQ ID NO: 210
SEQ ID NO: 211 QFISSY ATS QQSYNTLT PSA-2G7 HCDR SEQ ID NO: 216 SEQ
ID NO: 217 SEQ ID NO: 218 GDSVSSGSYY IYYSGST ARERCYYGSGRAP RCVWFDP
LCDR SEQ ID NO: 219 SEQ ID NO: 220 SEQ ID NO: 221 SSDVGGYNY DVS
SSYTSSSTLVV P5A-3C8 HCDR SEQ ID NO: 226 SEQ ID NO: 227 SEQ ID NO:
228 GFTVSSNY IYSGGST ARDLQEHGMDV LCDR SEQ ID NO: 229 SEQ ID NO: 230
SEQ ID NO: 231 QGISSY AAS QHLNSYPPGYT P5A-1D2 HCDR SEQ ID NO: 236
SEQ ID NO: 237 SEQ ID NO: 238 GFIVSSNY IYSGGST ARALQVGATSDYF DY
LCDR SEQ ID NO: 239 SEQ ID NO: 240 SEQ ID NO: 241 SSNIGAGYD GNS
QSCDSSLSVVV P5A-2F11 HCDR SEQ ID NO: 246 SEQ ID NO: 247 SEQ ID NO:
248 GYTFTSYD MNPNSGNT ARYIVVVPAAKGF DP LCDR SEQ ID NO: 249 SEQ ID
NO: 250 SEQ ID NO: 251 QSVLYSSNNKNY WAS QQYYSTPLT P5A-2E1 HCDR SEQ
ID NO: 256 SEQ ID NO: 257 SEQ ID NO: 258 GYSFTSYW IYPGDSDT
AQTSVTRNWFDP LCDR SEQ ID NO: 259 SEQ ID NO: 260 SEQ ID NO: 261
NIGSKS YDS QVWDSSSDHVV P5A-1C8 HCDR SEQ ID NO: 266 SEQ ID NO: 267
SEQ ID NO: 268 GYTFTSYY INPSGGST ARSARDYYDSSGY YYRAEYFQH LCDR SEQ
ID NO: 269 SEQ ID NO: 270 SEQ ID NO: 271 QDISNY DAS QQYDNLPSIT
P1A-1C10 HCDR SEQ ID NO: 276 SEQ ID NO: 277 SEQ ID NO: 278 GGTSSFYD
IIPRLDIA ARGRPGSEWAYGP FDL LCDR SEQ ID NO: 279 SEQ ID NO: 280 SEQ
ID NO: 281 QSSRAW KAS HQYNSSPFT P4A-1H6 HCDR SEQ ID NO: 286 SEQ ID
NO: 287 SEQ ID NO: 288 GFTFSSYG ISDDGSNQ AKRGGYCSTTSCLL RWVYFDF
LCDR SEQ ID NO: 289 SEQ ID NO: 290 SEQ ID NO: 291 QSISSY AAS
QQSYNTPT P4B-1F4 HCDR SEQ ID NO: 296 SEQ ID NO: 297 SEQ ID NO: 298
GFTFSSYG ISYDGSNK AKGPRYSSSWYISL YYYYGMDV LCDR SEQ ID NO: 299 SEQ
ID NO: 300 SEQ ID NO: 301 QSLVYSDGNTY KVS MQATHWPLYT P5A-1B6 HCDR
SEQ ID NO: 306 SEQ ID NO: 307 SEQ ID NO: 308 GFTFSSYA ISYDGSNK
ARDGQAITMVQGV IGPPFDY LCDR SEQ ID NO: 309 SEQ ID NO: 310 SEQ ID NO:
311 QDISNY DAS QQYDNLPYT P5A-1B8 HCDR SEQ ID NO: 316 SEQ ID NO: 317
SEQ ID NO: 318 GFTVSSNY IYPGGST ARETLAFDY LCDR SEQ ID NO: 319 SEQ
ID NO: 320 SEQ ID NO: 321 QGISSY AAS QQLNSYPPA P5A-1B9 HCDR SEQ ID
NO: 326 SEQ ID NO: 327 SEQ ID NO: 328 GGSISSYY ISYSGST
ASNGQYYDILTGQP PDYWYFDL LCDR SEQ ID NO: 329 SEQ ID NO: 330 SEQ ID
NO: 331 QSVLYSSNNKNY WAS QQYYSTPLT P5A-1D1 HCDR SEQ ID NO: 336 SEQ
ID NO: 337 SEQ ID NO: 338 GLTVSSNY IYSGGST ARDLYYYGMDV LCDR SEQ ID
NO: 339 SEQ ID NO: 340 SEQ ID NO: 341 QGISSY AAS QQLNSYPT P5A-1D10
HCDR SEQ ID NO: 346 SEQ ID NO: 347 SEQ ID NO: 348 QFTFSDYS ISQSGSTI
ARGVSPSYVWGSY RSLYHFDY LCDR SEQ ID NO: 349 SEQ ID NO: 350 SEQ ID
NO: 351 SSDVGGYNY DVS SSFTSSTTVVV P5A-2D11 HCDR SEQ ID NO: 356 SEQ
ID NO: 357 SEQ ID NO: 358 GYSFTSYW IYPGDSDT ARRDSTYGGNTDY LCDR SEQ
ID NO: 359 SEQ ID NO: 360 SEQ ID NO: 361 SSNIGSNT SNN AAWDDSLNGVV
P5A-2G9 HCDR SEQ ID NO: 366 SEQ ID NO: 367 SEQ ID NO: 368 GFTFSSYG
IWYDGSNK ARWFHTGGYFDY LCDR SEQ ID NO: 369 SEQ ID NO: 370 SEQ ID NO:
371 SDINVSSYN YYSDSDK MIWPSNALYV P5A-2H3 HCDR SEQ ID NO: 376 SEQ ID
NO: 377 SEQ ID NO: 378 GYSFTSYW IYPGDSDT ARRDSTYGGNTDY LCDR SEQ ID
NO: 379 SEQ ID NO: 380 SEQ ID NO: 381 SSNIGSNT SNN AAWDDSLNGVV
P5A-3A1 HCDR SEQ ID NO: 386 SEQ ID NO: 387 SEQ ID NO: 388 GFTVSSNY
IYSGGST ARDYGDFYFDY LCDR SEQ ID NO: 389 SEQ ID NO: 390 SEQ ID NO:
391 QSVSSSY GAS QQYGSSPRT P5A-3A6 HCDR SEQ ID NO: 396 SEQ ID NO:
397 SEQ ID NO: 398 GFTFDDYA ISWNSGTI AGGGTMVRGVIAG GGTHPVDDYYGM DV
LCDR SEQ ID NO: 399 SEQ ID NO: 400 SEQ ID NO: 401 SSDVGGYNY DVS
SSYTSSSTVV P5A-3B4 HCDR SEQ ID NO: 406 SEQ ID NO: 407 SEQ ID NO:
408 GYSFTSYW IYPGDSDT ARRDSTYGGNTDY LCDR SEQ ID NO: 409 SEQ ID NO:
410 SEQ ID NO: 411 SSNIGSNT SNN AAWDDSLNGVV P5A-3C12 HCDR SEQ ID
NO: 416 SEQ ID NO: 417 SEQ ID NO: 418 GFSLSTSGVG IYWDDDK
AHSLFLTVGYSSSW SPFDY LCDR SEQ ID NO: 419 SEQ ID NO: 420 SEQ ID NO:
421 QSVLYSSNNKNY WAS QQYYSTPHT P22A-1D1 HCDR SEQ ID NO: 426 SEQ ID
NO: 427 SEQ ID NO: 428 GFTVSSNY IYSGGST ARDRDYYGMDV LCDR SEQ ID NO:
429 SEQ ID NO: 430 SEQ ID NO: 431 QGISSY AAS LHLNSYRT
[0354] Table 2 below shows the heavy chain and light chain variable
region amino acid sequences of antibodies P2A-1A8, P2A-1A9,
P2B-2G11, P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4, P2C-1A3, P2C-1C8,
P2C-1C10, P2C-1D5, P2C-1F11, P2B-1G5, P2B-1A1, P2C- 1D7, P2B-1A10,
P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2,
P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6,
P5A-1B8, P5A- 1B9, P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9, P5A-2H3,
P5A-3A1, P5A-3A6, P5A-3B4, P5A-3C12, and P22A-1D1, and the
corresponding nucleic acid encoding sequence are shown in Table
3.
TABLE-US-00003 TABLE 2 Variable region amino acid sequences of 42
antibodies VH VL P2A-1A8 SEQ ID NO: 7 SEQ ID NO: 8
EVQLVESGGDLVQPGRSLRLSCA QSALTQPASVSGSPGQSITISCTG
ASGFAFDDYAMHWVRQAPGKG TSSDVGSYNLVSWYQQHPGKVP LEWVSGSTWNSGTIAYADSVKG
KLLIYDVNKRPSGISNRFSGSKS RFTISRDNAKKSLYLQMNSLRTE
GNTASLTISGLQAEDEADYYCRS DTALYYCAKLGGYSDYDYPRPG YTDSNTYVFGTGTKVTVL
DHYYGLDVWGQGTTVTVSS P2A-1A9 SEQ ID NO: 17 SEQ ID NO: 18
EVQLVESGGGLVQPGRSLRLSCA QSVLTQPPSVSGAPGQRVTISCT
ASGFTFDDYAMHWVRQVPGKGL GSSSNIGAGYDVHWYQQLPGTA
EWVSGISWNGGIIGYADSVKGRF PKLLIYGNNNRPSGVPDRFSGSK
TISRDNAKTSLYLQMNSLRAEDT SGTSASLAITGLQAEDEADYYCQ
ALYYCAKVAGRGDYDYYYGMD SYDSSLSGSVFGGGTKLTVL VWGQGTTVTVSS P2A-1A10
SEQ ID NO: 27 SEQ ID NO: 28 QVQLVQSGAEVKKPGASVKVSC
DIVMTQTPLSLPVTPGEPASISCR KASGYTFTGYYMHWVRQAPGQ
SSQSLLDSDDGNTYLDWYLQKP GLEWMGRINPNSGGTNYAQKFQ
GQSPQLLIYTLSYRASGVPDRFS GRVTMTRDTSISTAYMELSRLRS
GSGSGTDFTLKISRVEAEDVGVY DDTAVYYCARVPYCSSTSCHRD
YCMQRIEFPLTFGGGTKVEIK WYFDLWGRGTLVTVSS P2A-1B3 SEQ ID NO: 37 SEQ ID
NO: 38 EVQLVESGGGLVQPGGSLRLSCV EIVLTQSPGTLSLSPGERATLSCR
ASGFSFNRYSMNWLRQTPRKGL ASQSVSNDYLAWYQQKPGQAP
EWLSYISASGNTIYYADSVRGRF RLLIYYASSRATGIPDRFSGSGSG
TTSRDNAKNTLYLQMNSLRDDD TDFTLTISRLEPGDSAVYYCQQY
TAVYFCARPAMVREGTYNWFDP GDSPPITFGQGTRLEIK WGQGTLVTVSS P2B-2F6 SEQ ID
NO: 47 SEQ ID NO: 48 QVQLQESGPGLVKPSETLSLTCT
QSALTQPPSASGSPGQSVTISCTG VSGYSISSGYWGWIRQPPGKGL
TSSDVGGYNYVSWYQQHPGKA EWIGSIYHSGSTYYNPSLKTRVTI
PKLMIYEVSKRPSGVPDRFSGSK SVDTSKNQFSLKLSSVTAADTAV
SGNTASLTVSGLQAEDEADYYC YYCARAVVGIVVVPAAGRRAFDI SSYAGSNNLVFGGGTKLTVL
WGQGTMVTVSS P2B-2G4 SEQ ID NO: 57 SEQ ID NO: 58
QVQLVESGGGVVQPGRSLRLSCA QSALTQPRSVSGSPGQSVTISCT
ASGFTFSSYGIVIHWVRQAPGKGL GTSSDVGGYNYVSWYQQHPGK
EWVAVIWYDGSNKYYADSVKG APKLMIYDVSKRPSGVPDRFSGS
RFTISRDNSKNTLYLQMNSLRAE KSGNTASLTISGLQAEDEADYYC
DTAVYYCARGAAMVWLDYWG CSYAGSYTFVVFGGGTKLTVL QGTLVTVSS P2B-2G11 SEQ
ID NO: 61 SEQ ID NO: 62 EVQLVESGGGLVQPGRSLRLSCA
QSVLTQPPSVSGAPGQRVTISCT ASGFTFDDYAMHWVRQAPGKGL
GSSSNIGAGYDVHWYQQLPGTA EWVSGISWNGGIIGYADSVKGRF
PKLLIYGNNNRPSGVPDRFSGSK TISRDNAKTSLYLQMNSLKPEDT
SGTSASLAITGLQAEDEADYYCQ ALYYCAKVAGRGDYDYYYGMD SYDSSLSGSVFGGGTKLTVL
VWGQGTTVTVSS P2C-1A3 SEQ ID NO: 71 SEQ ID NO: 72
QVQLVESGGGLVKPGGSLRLSCA DIQLTQSPSFLSASVGDRVTITCR
ASGFTFSDYYMSWIRQAPGKGLE ASQGISSYLAWYQQKPGKAPKL
WVSYISSSGSTIYYADSVKGRFTI LIYAASTLQSGVPSRFSGSGSGTE
SRDNAKNSLYLQMNSLRAEDTA FTLTISSLQPEDFATYYCQQLNS
VYYCARDFSHQQLVPSWGQGTL YPLTFGGGTKVEIK VTVSS P2C-1C8 SEQ ID NO: 81
SEQ ID NO: 82 QVQLVESGGGVVQPGRSLRLSCA DVVMTQSPLSLPVTLGQPASISC
ASGFTFRSYGMHWVRQAPGKGL RSSQSLVYSDGNTYLNWFQQRP EWVAVIWYDGSNKYYADSVKG
GQSPRRLIYKVSIWDSGVPDRFS RFTISRDNSKNTLYLQMNSLRAE
GSGSGTDFTLKISRVEAEDVGVY DTAVYYCARDIEIVVVNIDYWGQ
YCMQGTHWPYTFGQGTKLEIK GTLVTVSS P2C-1C10 SEQ ID NO: 91 SEQ ID NO: 92
QVQLVQSGAEVKKPGSSVKVSC EIVLTQSPATLSLSPGERATLSCR
KASGGTFSSYAIIWVRQAPGQGL ASQSVSSYLAWYQQKPGQAPRL
EWMGGIIPIFGTANYAQKFQGRV LIYDASNRATGIPARFSGSGSGT
TITADESTSTAYMELSSLRSEDTA DFTLTISSLEPEDFAVYYCQQRS
VYYCARVVTGYYFDYWGQGTL NWPSFGQGTKLEIK VTVSS P2C-1D5 SEQ ID NO: 101
SEQ ID NO: 102 EVQLVESGGGLVQPGGSLRLSCA SYVLTQPPSVSVAPGKTARITCG
ASGFTFSSFAMSWVRQAPGKGLE GNNIGSKSVHWYQQKPGQAPVL
WVSAISGSGGSTYYADSVKGRFT VIYYDSDRPSGIPERFSGSNSGNT
ISRDNSKNTLYLQMNSLRAEDTA ATLTISRVEAGDEADYYCQVWD
VYYCAKDPDGSGSWYFDYWGQ SSSDHHVFGTGTKVTVL GTLVTVSS P2C-1F11 SEQ ID
NO: 111 SEQ ID NO: 112 EIVLTQSPGTLSLSPGERATLSCR
ASQSVSSSYLAWYQQKPGQAPR LLIYGASSRATGIPDRFSGSGSGT
DFTLTISRLEPEDFAVYYCQQYG SSPTFGQGTKLEIK P2B-1G5 SEQ ID NO: 142 SEQ
ID NO: 143 QVQLVQSGSELKKPGASVKVSC SYVLTQPPSVSVAPGKTARITCG
KASGYTFTTYVMNWVRQAPGQ GNNIGSKSVHWYQQKPGQAPVL GLEWMGWINTNTGNPTYAQGFT
VIYYDSDRPSGIPERFSGSNSGNT GRFVFSLDTSVSTASLQISSLKAE
ATLTISGVEAGDEADYYCQVWD DTAVYYCSCEITTLGGMDVWGQ SISDHRVFGGGTKLTVL
GTTVTVSS P2B-1A1 SEQ ID NO: 152 SEQ ID NO: 153
QVQLQESGPGLVKPSETLSLTCT QSALTQPASVSGSPGQSITISC
VSGGSISSYYWSWIRQPPGKGLE TGTSSDVGGYNYVSWYQQHP
WIGYIYYSGSTNYNPSLKSRVTIS GKAPKFMIYDVSKRPSGVSNR
VDTSKKQFSLKLSSVTAADTAVY FSGSKSGNTASLTISGLQAEDE
YCARLERDWPLDAFDIWGQGTM ADYYCSSYTSNNTFAFGGGT VTVSS KLTVL P2C-1D7 SEQ
ID NO: 162 SEQ ID NO: 163 EVQLVESGGGLIQPGGSLRLSCA
DVVMTQSPLSLPVTLGQPASISC ASGFTVSSNYMSWVRQAPGKGL
RSSQSLVYSDGNTYLNWFQQRP EWVSVIYSGGSTYYADSVKGRFT
GQSPRRLIYKVSNWDSGVPDRFS ISRDNSKNTLYLQMNSLRAEDTA
GSGSGTDFTLKISRVEAEDVGVY VYYCARELYEVGATDYWGQGTL
YCMQRYTLAGVFGPGTKVDIK VTVSS P2B-1A10 SEQ ID NO: 172 SEQ ID NO: 173
EVQLVESGGGLIQPGGSLRLSCA DIQMTQSPSSLSASVGDRVTITC
ASGFTVSSNYMSWVRQAPGKGL QASQDISNYFNWYQQKPGKAPK
EWVSVIYSGGSTYYADSVKGRFT LLIYDASNLETGVPSRFSGSGSG
ISRDNSKNTLYLQMNSLRAEDTA TDFTFTISSLQPEDIATYYCQQYD
VYYCAREGPKSITGTAFDIWGQG NLPMYTFGQGTKLEIK TIVTVSS P2B-1D9 SEQ ID NO:
182 SEQ ID NO: 183 QITLKESGPTLVKPTQTLTLTCTF QSVLTQPPSASGTPGQRVTISCS
SGFSLSTSGVGVGWIRQPPGKAL GSSSNIGSNYVYWYQQLPGTAP
EWLALIYWDDDKYYSPSLKSRLT KLLIYSNNQRPSGVPDRFSGSKS
ITKDTSKNQVVLTMTNMDPVDT GTSASLAISGLRSEDEADYYCAA
ATYYCAHTRILYYGSGSYYDYW WDDSLSGVVFGGGTKLTVL GQGTLVTVSS P2B-1E4 SEQ
ID NO: 192 SEQ ID NO: 193 QITLKESGPTLVKPTQTLTLTCTF
QSALTQPASVSGSPGQSITISCTG SGFSLSTSGVGVGWIRQPPGKAL
TSSDVGGYNYVSWYQQHPGKA EWLALIYWDDDKRYSPSLKSRLT
PKLMIYDVSKRPSGVSNRFSGSK ITKDTSKNQVVLTMTNMDPVDT
SGNTASLTISGLQAEDEADYYCS ATYYCAHQIVATIIDYWGQGTLV SYTSSSVVFGGGTKLTVL
TVSS P2B-1G1 SEQ ID NO: 202 SEQ ID NO: 203 EVQLVESGGGLVQPGGSLRLSCA
EIVLTQSPGTLSLSPGERATLSCR ASGFTVSSNYMSWVRQAPGKGL
ASQSVSSSYLAWYQQKPGQAPR EWVSVIYSGGSTYYADSVKGRFT
LLIYGASSRATGIPDRFSGSGSGT ISRDNSKNTLYLQMNSLRAEDTA
DFTLTISRLEPEDFAVYYCQQYG VYYCARDYGDYWFDPWGQGTL SSPRTFGQGTKLEIK VTVSS
P4A-2D9 SEQ ID NO: 212 SEQ ID NO: 213 QVQLVESGGGVVQPGRSLRLSCA
DIQMTQSPSSLSASVGDRVTITC ASGFTFSSYGIVIHWVRQSPGKGL
RASQFISSYLNWYQQKPGKAPK EWVAVISDDGSNQYYADSVKGR
LLIYATSILQTGVPSRFSGSGSGT FTISRDNSKNTLYLEINSLRVEDT
DFTLTISSLQPEDFATYYCQQSY AVYYCAKRGGYCSTTSCLVRWV NTLTFGPGTKVDIK
YFDYWGQGTLVTVSS P5A-2G7 SEQ ID NO: 222 SEQ ID NO: 223
QVQLQESGPGLVKPSETLSLTCT QSALTQPASVSGSPGQSITISCTG
VSGDSVSSGSYYWSWIRQPPGKG TSSDVGGYNYVSWYQQHPGKA
LEWIGYIYYSGSTNYNPSLKSRVT PKLMIYDVSNRPSGVSNRFSGSK
ISVDTSKNQFSLKLSSVTAADTA SGNTASLTISGLQAEDEADYYCS
VYYCARERCYYGSGRAPRCVWF SYTSSSTLVVFGGGTKLTVL DPWGQGTLVTVSS P5A-3C8
SEQ ID NO: 232 SEQ ID NO: 233 DIQLTQSPSSLSASVGDRVTITCR
ASQGISSYLAWYQQKPGKAPKL LIYAASTLQSGVPSRFSGSGSGT
DFTLTISSLQPEDFATYYCQHLN SYPPGYTFGQGTKLEIK P5A-1D2 SEQ ID NO: 242
SEQ ID NO: 243 QSVLTQPPSVSGAPGQRVTISCT GSSSNIGAGYDVHWYQQLPGTA
SPKLLIYGNSNRPSGVPDRFSGSK SGTSASLAITGLQAEDETDYYCQ
SCDSSLSVVVFGGGTKLTVL P5A-2F11 SEQ ID NO: 252 SEQ ID NO: 253
QVQLVQSGAEVKKPGASVKVSC DIVMTQSPDSLAVSLGERATINC
KASGYTFTSYDINWVRQATGQG KSSQSVLYSSNNKNYLAWYQQ LEWMGWMNPNSGNTGYAQKFQ
KPGQPPKLLIYWASTRESGVPDR GRVTMTRNTSISTAYMELSSLRS
FSGSGSGTDFTLTISSLQAEDVA EDTAVYYCARYIVVVPAAKGFD
VYYCQQYYSTPLTFGGGTKVEI PWGQGTLVTVSS K P5A-2E1 SEQ ID NO: 262 SEQ ID
NO: 263 EVQLVQSGAEVKKPGESLKISCK SYVLTQPPSVSVAPGKTARITCG
GSGYSFTSYWIGWVRQMPGKGL GNNIGSKSVHWYQQKPGQAPVL
EWMGIIYPGDSDTRYSPSFQGQV VIYYDSDRPSGIPERFSGSNSGNT
TISADKSISTAYLQWSSLKASDTA ATLTISRVEAGDEADYYCQVWD
MYYCAQTSVTRNWFDPWGQGT SSSDHVVFGGGTKLTVL LVTVSS P5A-1C8 SEQ ID NO:
272 SEQ ID NO: 273 QVQLVQSGAEVKKPGASVKVSC DIQMTQSPSSLSASVGDRVTITC
KASGYTFTSYYMHWVRQAPGQG QASQDISNYLNWYQQKPGKAPK
LEWMGIINPSGGSTSYAQKFQGR LLIYDASNLETGVPSRFSGSGSG
VTMTRDTSTSTVYMELSSLRSED TDFTFTISSLQPEDIATYYCQQYD
TAVYYCARSARDYYDSSGYYYR NLPSITFGQGTRLEIK AEYFQHWGQGTLVTVSS P1A-1C10
SEQ ID NO: 282 SEQ ID NO: 283 QVQLVQSGAEVKNPGSSVKVSC
DIQMTQSPSTLSASVGDRVTITC KAGGGTSSFYDINWVRQAPGQG
RASQSSRAWLAWYQQKPGKAP LEWIGKIIPRLDIADYAQKSQGRV
KLLISKASSLESGVPSRFSGSGYG TITADKSTSTVYLELSSLKSDDTA
TEFTLTISSLQPDDSATYYCHQY VYFCARGRPGSEWAYGPFDLWG NSSPFTFGPGTKVQIK
QGTLVTVSS P4A-1H6 SEQ ID NO: 292 SEQ ID NO: 293
QVQLVESGGGVVQPGRSLRLSCA DIQMTQSPSSLSASVGDRVTITC
ASGFTFSSYGIVIHWVRQSPGKGL RASQSISSYLHWYQQKPGKAPN
EWVAVISDDGSNQYYADSVKGR LLIYAASSLQSGVPSRFSGSGSGT
FTISRDNSKNTLYLQMNSLRVED DFTLTISSLQPEDFATYYCQQSY
TAVYYCAKRGGYCSTTSCLLRW NTPTFGPGTKVDIK VYFDFWGQGTLATVS S P4B-1F4 SEQ
ID NO: 302 SEQ ID NO: 303 QVQLVESGGGVVQPGRSLRLSCA
DVVMTQSPLSLPVTLGQPASISC ASGFTFSSYGIVIHWVRQAPGKGL
RSSQSLVYSDGNTYLNWFQQRP EWVAVISYDGSNKYYADSVKGR
GQSPRRLIYKVSNRDSGVPDRFS FTISRDNSKNTLYLQINSLRAEDT
GSGSGTDFTLKISRVEAEDVGVY AVYYCAKGPRYSSSWYISLYYY
YCMQATHWPLYTFGQGTKLEIK YGMDVWGQGTTVTVSS P5A-1B6 SEQ ID NO: 312 SEQ
ID NO: 313 QVQLVESGGGVVQPGRSLRLSCA DIQMTQSPSSLSASVGDRVTITC
ASGFTFSSYAMIHWVRQAPGKGL QASQDISNYLNWYQQKPGKAPK
EWVAVISYDGSNKYYADSVKGR LLIYDASNLETGVPSRFSGSGSG
FTISRDNSKNTLYLQMNSLRAED TDFTFTISSLQPEDIATYYCQQYD
TAVYYCARDGQAITMVQGVIGPP NLPYTFGQGTKLEIK FDYWGQGTLVTVSS P5A-1B8 SEQ
ID NO: 322 SEQ ID NO: 323 EVQLVESGGGLIQPGGSLRLSCA
DIQLTQSPSFLSASVGDRVTITCR ASGFTVSSNYMSWVRQAPGKGL
ASQGISSYLAWYQQKPGKAPKL EWVSVIYPGGSTFYADSVKGRFT
LIYAASTLQSGVPSRFSGSGSGTE ISRDNSKNTLYLQMNSLRAEDTA
FTLTISSLQPEDFATYYCQQLNS
VYYCARETLAFDYWGQGTLVTV YPPAFGGGTKVEIK SS P5A-1B9 SEQ ID NO: 332 SEQ
ID NO: 333 QVQLQESGPGLVKPSETLSLTCT DIVMTQSPDSLAVSLGERATINC
VSGGSISSYYWSWIRQPPGKGLE KSSQSVLYSSNNKNYLAWYQQ
WIGYISYSGSTNYNPSLKSRVTIS KPGQPPKLLIYWASTRESGVPDR
LDTSKNQFSLKLSSVTAADTAVY FSGSGSGTDFTLTISSLQAEDVA
YCASNGQYYDILTGQPPDYWYF VYYCQQYYSTPLTFGGGTKVEI DLWGRGTLVTVSS K
P5A-1D1 SEQ ID NO: 342 SEQ ID NO: 343 EVQLVESGGGLIQPGGSLRLSCA
DIQLTQSPSFLSASVGDRVTITCR ASGLTVSSNYMSWVRQAPGKGL
ASQGISSYLAWYQQKPGKAPKL EWVSVIYSGGSTYYADSVKGRFT
LIYAASTLQSGVPSRFSGSGSGT ISRDNSKNTLYLQMNSLRAEDTA
DFTLTISSLQPEDFATYYCQQLN VYYCARDLYYYGMDVWGQGTT SYPTFGQGTRLEIK VTVST
P5A-1D10 SEQ ID NO: 352 SEQ ID NO: 353 QVQLVESGGGLVKPGGSLRLSCA
QSALTQPASVSGSPGQSITISCTG ASQFTFSDYSMTWIRQAPGKGLE
TSSDVGGYNYVSWYQQHPGKA WVSYISQSGSTIYYADSVKGRFTI
PKLMIYDVSNRPSGVSNRFSASK SRDNAKNSLYLQMNSLRAEDTA
SGNTASLTISGLQAEDEADYYCS VYYCARGVSPSYVWGSYRSLYH SFTSSTTVVVFGGGTKLTVL
FDYWGQGTLVTVSS P5A-2D11 SEQ ID NO: 362 SEQ ID NO: 363
EVQLVQSGAEVKKPGESLKISCK QSVLTQPPSASGTPGQRVTISCS
GSGYSFTSWIGWVRQMPGKGL GSSSNIGSNTVNWYQQLPGTAP
EWMGIIYPGDSDTRYSPSFQGQV KLLIYSNNQRPSGVPDRFSGSKS
TISADKSISTAYLQWSSLKASDTA GTSASLAISGLQSEDEADYYCAA
MYYCARRDSTYGGNTDWGQG WDDSLNGVVFGGGTKLTVL TLVTVSS P5A-2G9 SEQ ID NO:
372 SEQ ID NO: 373 QVQLVESGGGVVQPGRSLRLSCA QPVLTQPPSSSASPGESARLTCTL
ASGFTFSSYGMEIWVRQAPGKGL PSDINVSSYNIYWYQQKPGSPPR
EWVAVIWYDGSNKYYADSVKG YLLYYYSDSDKGQGSGVPSRFS
RFTISRDNSKNTLYLQMNSLRAE GSKDASANTGILLISGLQSEDEA
DTAVYYCARWFHTGGYFDYWG DYYCMIWPSNALYVFGTGTKVT QGTLVTVSS VL P5A-2H3
SEQ ID NO: 382 SEQ ID NO: 383 EVQLVQSGAEVKKPGESLKISCK
QSVLTQPPSASGTPGQRVTISCS GSGYSFTSWIGWVRQMPGKGL
GSSSNIGSNTVNWYQQLPGTAP EWMGIIYPGDSDTRYSPSFQGQV
KLLIYSNNQRPSGVPDRFSGSKS TISAEKSISTAYLQWSSLKASDTA
GTSASLAISGLQSEDEADYYCAA MYYCARRDSTYGGNTDWGQG WDDSLNGVVFGGGTKLTVL
TLVTVSS P5A-3A1 SEQ ID NO: 392 SEQ ID NO: 393
EVQLVESGGGLIQPGGSLRLSCA EIVLTQSPGTLSLSPGERATLSCR
ASGFTVSSNYMSWVRQAPGKGL ASQSVSSSYLAWYQQKPGQAPR
EWVSVIYSGGSTYYADSVKGRFT LLIYGASSRATGIPDRFSGSGSGT
ISRDNSKNTLYLQMNSLRAEDTA DFTLTISRLEPEDFAVYYCQQYG
VYYCARDYGDFYFDYWGQGTL SSPRTFGQGTKLEIK VTVSS P5A-3A6 SEQ ID NO: 402
SEQ ID NO: 403 EVQLVESGGGLVQPGRSLRLSCA QSALTQPASVSGSPGQSITISCTG
ASGFTFDDYAMHWVRQAPGKGL TSSDVGGYNYVSWYQQHPGKA
EWVSGISWNSGTIGYADSVKGRF PKLMIYDVSNRPSGVSNRFSGSK
IISRDNAKNSLYLQMNSLRAEDT SGNTASLTISGLQAEDEADYYCS
ALYYCAGGGTMVRGVIAGGGTH SYTSSSTVVFGGGTKLTVL PVDDYYGMDVWGQGTTVTVSS
P5A-3B4 SEQ ID NO: 412 SEQ ID NO: 413 EVQLVQSGAEVKEPGESLKISCK
QSVLTQPPSASGTPGQRVTISCS GSGYSFTSYWIGWVRQMPGKGL
GSSSNIGSNTVNWYQQLPGTAP EWMGIIYPGDSDTRYSPSFQGQV
KLLIYSNNQRPSGVPDRFSGSKS TISADKSISTAYLQWSSLKASDTA
GTSASLAISGLQSEDEADYYCAA MYYCARRDSTYGGNTDYWGQG WDDSLNGVVFGGGTKLTVL
TLVTVSS P5A-3C12 SEQ ID NO: 422 SEQ ID NO: 423
QITLKESGPTLVKPTQTLTLTCTF DIVMTQSPDSLAVSLGERATINC
SGFSLSTSGVGVGWIRQPPGKAL KSSQSVLYSSNNKNYLAWYQQ
EWLALIYWDDDKRYSPSLKSRLT KPGQPPKLLIYWASTRESGVPDR
ITKDTSKNQVVLTMTNMDPVDT FSGSGSGTDFTLTISSLQAEDVA
ATYYCAHSLFLTVGYSSSWSPFD VYYCQQYYSTPHTFGQGTKLEI YWGQGTLVTVSS K
P22A-1D1 SEQ ID NO: 432 SEQ ID NO: 433 DIQLTQSPSFLSASVGDRVTITCR
ASQGISSYLAWYQQKPGKAPKL LIYAASTLQSGVPSRFSGSGSGTE
FTLTISSLQPEDFATYYCLHLNSY RTFGLGTKVEIK
TABLE-US-00004 TABLE 3 Variable region nucleotide sequences of 42
antibodies VHnu (nucleic acid) VLnu (nucleic acid) P2A-1A8 SEQ ID
NO: 9 SEQ ID NO: 10 GAAGTGCAGCTGGTGGAGTCTG CAGTCTGCCCTGACTCAGCCTG
GGGGAGACTTGGTACAGCCTGG CCTCCGTGTCTGGGTCTCCTGG
CAGGTCCCTGAGACTCTCCTGC ACAGTCGATCACCATCTCCTGC
GCAGCCTCTGGATTCGCCTTTG ACTGGAACCAGCAGTGATGTT ATGATTATGCCATGCACTGGGT
GGGAGTTATAACCTTGTCTCCT CCGGCAAGCTCCAGGGAAGGG GGTACCAACAGCACCCAGGCA
CCTGGAGTGGGTCTCAGGTAGT AAGTCCCCAAACTCTTGATTTA
ACTTGGAATAGTGGGACCATAG TGATGTCAATAAGCGGCCCTCA
CCTATGCGGACTCTGTGAAGGG GGGATTTCCAATCGCTTCTCTG
CCGATTCACCATCTCCAGAGAC GCTCCAAGTCTGGCAACACGG AACGCCAAGAAGTCCCTGTATC
CCTCCCTGACCATCTCTGGGCT TGCAAATGAACAGTCTGAGAAC CCAGGCTGAGGACGAGGCTGA
TGAGGACACGGCCTTATATTAC TTATTACTGCAGATCATATACA
TGTGCAAAGTTGGGGGGCTACA GACAGCAACACTTATGTCTTCG
GTGACTACGATTACCCGAGGCC GAACTGGGACCAAGGTCACCG GGGAGACCACTATTACGGTTTG
TCCTA GACGTCTGGGGCCAAGGGACCA CGGTCACCGTCTCCTCA P2A-1A9 SEQ ID NO:
19 SEQ ID NO: 20 GAAGTGCAGCTGGTGGAGTCTG CAGTCTGTGCTGACGCAGCCGC
GGGGAGGCTTGGTACAGCCTGG CCTCAGTGTCTGGGGCCCCAGG
CAGGTCCCTGAGACTCTCCTGT GCAGAGGGTCACCATCTCCTGC
GCAGCCTCTGGATTCACCTTTG ACTGGGAGCAGCTCCAACATC ATGATTATGCCATGCACTGGGT
GGGGCAGGTTATGATGTACAC CCGGCAAGTTCCAGGGAAGGGC TGGTACCAGCAACTTCCAGGA
CTGGAGTGGGTCTCAGGTATTA ACAGCCCCCAAACTCCTCATCT
GTTGGAATGGTGGTATCATAGG ATGGTAACAACAATCGCCCCTC
CTACGCGGACTCTGTGAAGGGC AGGGGTCCCTGACCGATTCTCT
CGATTCACCATCTCCAGAGACA GGCTCCAAGTCTGGCACCTCAG
ACGCCAAGACTTCCCTGTATCT CCTCCCTGGCCATCACTGGGCT
GCAAATGAACAGTCTGAGAGCT CCAGGCTGAGGATGAGGCTGA GAGGACACGGCCTTGTATTACT
TTATTACTGCCAGTCCTATGAC GTGCAAAAGTCGCGGGAAGGG AGCAGCCTGAGTGGTTCGGTAT
GGGATTACGACTATTACTATGG TCGGCGGAGGGACCAAGCTGA TATGGACGTCTGGGGCCAAGGG
CCGTCCTA ACCACGGTCACCGTCTCCTCA P2A-1A10 SEQ ID NO: 29 SEQ ID NO: 30
CAGGTGCAGCTGGTGCAGTCTG GATATTGTGATGACCCAGACTC GGGCTGAGGTGAAGAAGCCTG
CACTCTCCCTGCCCGTCACCCC GGGCCTCAGTGAAGGTCTCCTG
TGGAGAGCCGGCCTCCATCTCC CAAGGCTTCTGGATACACCTTC
TGCAGGTCTAGTCAGAGCCTCT ACCGGCTACTATATGCACTGGG TGGATAGTGATGATGGAAACA
TGCGACAGGCCCCTGGACAAGG CCTATTTGGACTGGTACCTGCA
GCTTGAGTGGATGGGACGGATC GAAGCCAGGGCAGTCTCCACA AACCCTAACAGTGGTGGCACAA
GCTCCTGATCTATACGCTTTCC ACTATGCACAGAAGTTTCAGGG
TATCGGGCCTCTGGAGTCCCAG CAGGGTCACCATGACCAGGGAC ACAGGTTCAGTGGCAGTGGGT
ACGTCCATCAGCACAGCCTACA CAGGCACTGATTTCACACTGAA
TGGAGCTGAGCAGGCTGAGATC AATCAGCAGGGTGGAGGCTGA TGACGACACGGCCGTGTATTAC
GGATGTTGGAGTTTATTACTGC TGTGCGAGAGTCCCCTATTGTA
ATGCAACGTATAGAGTTTCCGC GTAGTACCAGCTGCCATCGGGA TCACTTTCGGCGGAGGGACCA
CTGGTACTTCGATCTCTGGGGC AGGTGGAGATCAAA CGTGGCACCCTGGTCACTGTCT CCTCA
P2A-1B3 SEQ ID NO: 39 SEQ ID NO: 40 GAGGTGCAGCTGGTGGAGTCTG
GAAATTGTGTTGACGCAGTCTC GGGGAGGCTTGGTACAGCCTGG
CAGGCACCCTGTCTTTGTCTCC GGGGTCCCTCAGACTCTCCTGT AGGGGAAAGAGCCACCCTCTC
GTCGCCTCTGGATTCTCCTTCAA CTGCAGGGCCAGTCAGAGTGT
TCGATATAGTATGAATTGGCTC TAGCAACGACTACTTAGCCTGG
CGCCAGACTCCACGGAAGGGGC TACCAGCAGAAACCTGGCCAG TGGAGTGGCTTTCATACATCAG
GCTCCCAGGCTCCTCATCTACT TGCCAGTGGAAACACCATATAC ATGCATCCAGCAGGGCCACTG
TACGCTGACTCTGTGAGGGGCC GCATCCCAGACAGGTTCAGTG GATTCACCACCTCCAGAGACAA
GCAGTGGGTCTGGGACAGACT TGCCAAGAACACACTGTATCTG TCACTCTCACCATCAGCAGACT
CAAATGAACAGCCTGCGAGACG GGAGCCTGGAGATTCTGCAGT ACGACACGGCTGTCTATTTCTG
GTATTACTGTCAGCAGTATGGT TGCGCGACCCGCTATGGTTCGG
GACTCACCTCCGATCACCTTCG GAGGGGACCTACAACTGGTTCG GCCAAGGGACACGACTGGAGA
ACCCCTGGGGCCAGGGAACCCT TTAAA GGTCACCGTCTCCTCA P2B-2F6 SEQ ID NO: 49
SEQ ID NO: 50 CAGGTGCAGCTGCAGGAGTCGG CAGTCTGCCCTGACTCAGCCTC
GCCCAGGACTGGTGAAGCCTTC CCTCCGCGTCCGGGTCTCCTGG
GGAGACCCTGTCCCTCACCTGC ACAGTCAGTCACCATCTCCTGC
ACTGTCTCTGGTTACTCCATCAG ACTGGAACCAGCAGTGACGTT
CAGTGGTTACTACTGGGGCTGG GGTGGTTATAACTATGTCTCCT
ATCCGGCAGCCCCCAGGGAAGG GGTACCAACAGCACCCAGGCA GGCTGGAGTGGATTGGGAGTAT
AAGCCCCCAAACTCATGATTTA CTATCATAGTGGGAGCACCTAC TGAGGTCAGTAAGCGGCCCTC
TACAACCCGTCCCTCAAGACTC AGGGGTCCCTGATCGCTTCTCT
GAGTCACCATATCAGTAGACAC GGCTCCAAGTCTGGCAACACG GTCCAAGAACCAGTTCTCCCTG
GCCTCCCTGACCGTCTCTGGGC AAGCTGAGCTCTGTGACCGCCG TCCAGGCTGAGGATGAGGCTG
CAGACACGGCCGTCTATTACTG ATTATTACTGCAGCTCATATGC
TGCGAGAGCGGTGGTAGGGATT AGGCAGCAACAATTTGGTGTTC
GTAGTAGTACCAGCTGCCGGTC GGCGGAGGGACCAAGCTGACC GTCGGGCTTTTGATATCTGGGG
GTCCTA CCAAGGGACAATGGTCACCGTC TCCTCA P2B-2G4 SEQ ID NO: 59 SEQ ID
NO: 60 CAGGTGCAGCTGGTGGAGTCTG CAGTCTGCCCTGACTCAGCCTC
GGGGAGGCGTGGTCCAGCCTGG GCTCAGTGTCCGGGTCTCCTGG
GAGGTCCCTGAGACTCTCCTGT ACAGTCAGTCACCATCTCCTGC
GCAGCGTCTGGATTCACCTTCA ACTGGAACCAGCAGTGATGTT GTAGCTATGGCATGCACTGGGT
GGTGGTTATAACTATGTCTCCT CCGCCAGGCTCCAGGCAAGGGG GGTACCAACAGCACCCAGGCA
CTGGAGTGGGTGGCAGTTATAT AAGCCCCCAAACTCATGATTTA
GGTATGATGGAAGTAATAAATA TGATGTCAGTAAGCGGCCCTCA
CTATGCAGACTCCGTGAAGGGC GGGGTCCCTGATCGCTTCTCTG
CGATTCACCATCTCCAGAGACA GCTCCAAGTCTGGCAACACGG ATTCCAAGAACACGCTGTATCT
CCTCCCTGACCATCTCTGGGCT GCAAATGAACAGCCTGAGAGCC CCAGGCTGAGGATGAGGCTGA
GAGGACACGGCTGTGTATTACT TTATTACTGCTGCTCATATGCA
GTGCGAGAGGGGCAGCTATGGT GGCAGCTACACTTTCGTGGTAT
TTGGCTTGACTACTGGGGCCAG TCGGCGGAGGGACCAAGCTGA GGAACCCTGGTCACCGTCTCCT
CCGTCCTA CA P2B-2G11 SEQ ID NO: 63 SEQ ID NO: 64
GAAGTGCAGCTGGTGGAGTCTG CAGTCTGTGCTGACGCAGCCGC
GGGGAGGCTTGGTACAGCCTGG CCTCAGTGTCTGGGGCCCCAGG
CAGGTCCCTGAGACTCTCCTGT GCAGAGGGTCACCATCTCCTGC
GCAGCCTCTGGATTCACCTTTG ACTGGGAGCAGCTCCAACATC ATGATTATGCCATGCACTGGGT
GGGGCAGGTTATGATGTACAC CCGGCAAGCTCCAGGGAAGGG TGGTACCAGCAACTTCCAGGA
CCTGGAGTGGGTCTCAGGTATT ACAGCCCCCAAACTCCTCATCT
AGTTGGAATGGTGGTATCATAG ATGGGAACAACAATCGGCCCT GCTATGCGGACTCTGTGAAGGG
CAGGGGTCCCTGACCGATTCTC CCGATTCACCATCTCCAGAGAC
TGGCTCCAAGTCTGGCACCTCA AACGCCAAGACTTCCCTGTATC
GCCTCCCTGGCCATCACTGGGC TGCAAATGAACAGTCTGAAACC TCCAGGCTGAGGATGAGGCTG
TGAGGACACGGCCTTGTATTAC ATTATTACTGCCAGTCCTATGA TGTGCAAAAGTCGCGGGAAGG
CAGCAGCCTGAGTGGTTCGGT GGGGATTACGACTACTACTACG ATTCGGCGGAGGGACCAAGCT
GTATGGACGTCTGGGGCCAAGG GACCGTCCTA GACCACGGTCACCGTCTCCTCA P2C-1A3
SEQ ID NO: 73 SEQ ID NO: 74 CAGGTGCAGCTGGTGGAGTCTG
GACATCCAGTTGACCCAGTCTC GGGGAGGCTTGGTCAAGCCTGG
CATCCTTCCTGTCTGCATCTGT AGGGTCCCTGAGACTCTCCTGT AGGAGACAGAGTCACCATCAC
GCAGCCTCTGGATTCACCTTCA TTGCCGGGCCAGTCAGGGCATT
GTGACTACTACATGAGCTGGAT AGCAGTTATTTAGCCTGGTATC CCGCCAGGCTCCAGGGAAGGG
AGCAAAAACCAGGGAAAGCCC GCTGGAGTGGGTTTCATACATT CTAAGCTCCTGATCTATGCTGC
AGTAGTAGTGGTAGTACCATAT ATCCACTTTGCAAAGTGGGGTC
ACTACGCAGACTCTGTGAAGGG CCATCAAGGTTCAGCGGCAGT CCGATTCACCATCTCCAGGGAC
GGATCTGGGACAGAATTCACT AACGCCAAGAACTCACTGTATC CTCACAATCAGCAGCCTGCAG
TGCAAATGAACAGCCTGAGAGC CCTGAAGATTTTGCAACTTATT
CGAGGACACGGCTGTGTATTAC ACTGTCAACAGCTTAATAGTTA
TGTGCGAGAGATTTTTCTCATC CCCGCTCACTTTCGGCGGAGGG
AGCAGCTGGTACCTTCCTGGGG ACCAAGGTGGAGATCAAA CCAGGGAACCCTGGTCACCGTC
TCCTCA P2C-1C8 SEQ ID NO: 83 SEQ ID NO: 84 CAGGTGCAGCTGGTGGAGTCTG
GATGTTGTGATGACTCAGTCTC GGGGAGGCGTGGTCCAGCCTGG
CACTCTCCCTGCCCGTCACCCT GAGGTCCCTGAGACTCTCCTGT
TGGACAGCCGGCCTCCATCTCC GCAGCGTCTGGATTCACCTTCA
TGCAGGTCTAGTCAAAGCCTCG GGAGCTATGGCATGCACTGGGT TATACAGTGATGGAAACACCT
CCGCCAGGCTCCAGGCAAGGGG ACTTGAATTGGTTTCAGCAGAG
CTGGAGTGGGTGGCAGTTATCT GCCAGGCCAATCTCCAAGGCG GGTATGATGGAAGTAATAAATA
CCTAATTTATAAGGTTTCTATC CTATGCAGACTCCGTGAAGGGC TGGGACTCTGGGGTCCCAGAC
CGATTCACCATCTCCAGAGACA AGATTCAGCGGCAGTGGGTCA ATTCCAAGAACACGCTGTATCT
GGCACTGATTTCACACTGAAA GCAAATGAACAGCCTGAGAGCC ATCAGCAGGGTGGAGGCTGAG
GAGGACACGGCTGTGTATTACT GATGTTGGGGTTTATTACTGCA
GTGCGAGAGATATAGAGATAGT TGCAAGGTACACACTGGCCGT AGTGGTAAATATTGACTACTGG
ACACTTTTGGCCAGGGGACCA GGCCAGGGAACCCTGGTCACCG AGCTGGAGATCAAA
TCTCCTCA P2C-1C10 SEQ ID NO: 93 SEQ ID NO: 94
CAGGTGCAGCTGGTGCAGTCTG GAAATTGTGTTGACACAGTCTC GGGCTGAGGTGAAGAAGCCTG
CAGCCACCCTGTCTTTGTCTCC GGTCCTCGGTGAAGGTCTCCTG AGGGGAAAGAGCCACCCTCTC
CAAGGCTTCTGGAGGCACCTTC CTGCAGGGCCAGTCAGAGTGT AGCAGCTATGCTATCATCTGGG
TAGCAGCTACTTAGCCTGGTAC TGCGACAGGCCCCTGGACAAGG CAACAGAAACCTGGCCAGGCT
GCTTGAGTGGATGGGAGGGATC CCCAGGCTCCTCATCTATGATG
ATCCCTATCTTTGGTACAGCAA CATCCAACAGGGCCACTGGCA ACTACGCACAGAAGTTCCAGGG
TCCCAGCCAGGTTCAGTGGCA CAGAGTCACGATTACCGCGGAC GTGGGTCTGGGACAGACTTCA
GAATCCACGAGCACAGCCTACA CTCTCACCATCAGCAGCCTAGA
TGGAGCTGAGCAGCCTGAGATC GCCTGAAGATTTTGCAGTTTAT
TGAGGACACGGCCGTGTATTAC TACTGTCAGCAGCGTAGCAACT
TGTGCGAGAGTGGTAACGGGGT GGCCTTCTTTTGGCCAGGGGAC
ACTACTTTGACTACTGGGGCCA CAAGCTGGAGATCAAA GGGAACCCTGGTCACCGTCTCC TCA
P2C-1D5 SEQ ID NO: 103 SEQ ID NO: 104 GAGGTGCAGCTGGTGGAGTCTG
TCCTATGTGCTGACTCAGCCAC GGGGAGGCTTGGTACAGCCTGG
CCTCAGTGTCAGTGGCCCCAGG GGGGTCCCTGAGACTCTCCTGT AAAGACGGCCAGGATTACCTG
GCAGCCTCTGGATTCACCTTTA TGGGGGAAACAACATTGGAAG GCAGCTTTGCCATGAGCTGGGT
TAAAAGTGTGCACTGGTACCA CCGCCAGGCTCCAGGGAAGGG GCAGAAGCCAGGCCAGGCCCC
GCTGGAGTGGGTCTCAGCTATT TGTGCTGGTCATCTATTATGAT
AGTGGTAGTGGTGGTAGCACAT AGCGACCGGCCCTCAGGGATC ACTACGCAGACTCCGTGAAGGG
CCTGAGCGATTCTCTGGCTCCA CCGGTTCACCATCTCCAGAGAC
ACTCTGGGAACACCGCCACCCT AATTCCAAGAACACGCTGTATT GACCATCAGCAGGGTCGAAGC
TGCAAATGAACAGCCTGAGAGC CGGGGATGAGGCCGACTATTA CGAGGACACGGCCGTATATTAC
CTGTCAGGTGTGGGATAGTAGT TGTGCGAAAGATCCGGATGGTT
AGTGATCATCATGTCTTCGGAA CGGGGAGTTGGTACTTTGACTA
CTGGGACCAAGGTCACCGTCCT CTGGGGCCAGGGAACCCTGGTC A ACCGTCTCCTCA
P2C-1F11 SEQ ID NO: 113 SEQ ID NO: 114 GAGGTGCAGCTGGTGGAGTCTG
GAAATTGTGTTGACGCAGTCTC GGGGAGGCTTGGTCCAGCCTGG
CAGGCACCCTGTCTTTGTCTCC GGGGTCCCTGAGACTCTCCTGT AGGGGAAAGAGCCACCCTCTC
GCAGCCTCTGGAATCACCGTCA CTGCAGGGCCAGTCAGAGTGT GTAGCAACTACATGAACTGGGT
TAGCAGCAGCTACTTAGCCTGG CCGCCAGGCTCCAGGGAAGGG TACCAGCAGAAACCTGGCCAG
GCTGGAGTGGGTCTCACTTATT GCTCCCAGGCTCCTCATCTATG
TATAGCGGTGGTAGCACATACT GTGCATCCAGCAGGGCCACTG ACGCAGACTCCGTGAAGGGCAG
GCATCCCAGACAGGTTCAGTG ATTCACCATCTCCAGAGACAAT GCAGTGGGTCTGGGACAGACT
TCCAAGAACACGTTGTATCTTC TCACTCTCACCATCAGCAGACT AAATGAACAGCCTGAGAGCCG
GGAGCCTGAAGATTTTGCAGT AGGACACGGCTGTGTATCACTG GTATTACTGTCAGCAGTATGGT
TGCGAGAGATCTGGTGGTATAC AGCTCACCCACTTTTGGCCAGG
GGTATGGACGTCTGGGGCCAAG GGACCAAGCTGGAGATCAAA GGACCACGGTCACCGTCTCCTC
A P2B-1G5 SEQ ID NO: 144 SEQ ID NO: 145 CAGGTGCAGCTGGTGCAATCTG
TCCTATGTGCTGACTCAGCCAC GGTCTGAGTTGAAGAAGCCTGG
CCTCAGTGTCAGTGGCCCCAGG GGCCTCAGTGAAGGTTTCCTGC AAAGACGGCCAGGATTACCTG
AAGGCTTCTGGATACACCTTCA TGGGGGAAACAACATTGGAAG CTACCTATGTTATGAATTGGGT
TAAAAGTGTGCACTGGTACCA GCGACAGGCCCCTGGACAAGG GCAGAAGCCAGGCCAGGCCCC
GCTTGAGTGGATGGGATGGATC TGTGCTGGTCATCTATTATGAT
AACACCAACACTGGGAACCCAA AGCGACCGGCCCTCAGGGATC CGTATGCCCAGGGCTTCACAGG
CCTGAGCGATTCTCTGGCTCCA ACGGTTTGTCTTCTCCTTGGACA
ACTCTGGGAACACGGCCACCC CCTCTGTCAGCACGGCATCTCT TGACCATCAGCGGGGTCGAAG
GCAGATCAGCAGCCTAAAGGCT CCGGGGATGAGGCCGACTATT GAGGACACTGCCGTGTATTACT
ACTGTCAGGTGTGGGATAGTAT
GTTCGTGTGAAATAACCACCTT TAGTGATCATCGGGTGTTCGGC
GGGCGGTATGGACGTCTGGGGC GGAGGGACCAAGCTGACCGTC CAAGGGACCACGGTCACCGTCT
CTA CCTCA P2B-1A1 SEQ ID NO: 154 SEQ ID NO: 155
CAGGTGCAGCTGCAGGAGTCGG CAGTCTGCCCTGACTCAGCCTG
GCCCAGGACTGGTGAAGCCTTC CCTCCGTGTCTGGGTCTCCTGG
GGAGACCCTGTCCCTCACCTGC ACAGTCGATCACCATCTCCTGC
ACTGTCTCTGGTGGCTCCATCA ACTGGAACCAGCAGTGACGTT GTAGTTACTACTGGAGCTGGAT
GGTGGTTATAACTATGTCTCCT CCGGCAGCCCCCAGGGAAGGG GGTACCAACAGCACCCAGGCA
ACTGGAGTGGATTGGGTATATC AAGCCCCCAAATTCATGATTTA
TATTACAGTGGGAGCACCAACT TGATGTCAGTAAGCGGCCCTCA
ACAACCCCTCCCTCAAGAGTCG GGGGTTTCTAATCGCTTCTCTG
AGTCACCATATCAGTAGACACG GCTCCAAGTCTGGCAACACGG TCCAAGAAGCAGTTCTCCCTGA
CCTCCCTGACCATCTCTGGGCT AGCTGAGCTCTGTGACCGCTGC CCAGGCTGAGGACGAGGCTGA
GGACACGGCCGTGTATTACTGT TTATTACTGCAGCTCATATACA
GCGAGGCTCGAACGAGACTGGC AGCAACAACACTTTCGCGTTCG
CACTTGATGCTTTTGATATCTGG GCGGAGGGACCAAGCTGACCG
GGCCAAGGGACAATGGTCACCG TCCTA TCTCCTCA P2C-1D7 SEQ ID NO: 164 SEQ ID
NO: 165 GAGGTGCAGCTGGTGGAGTCTG GATGTTGTGATGACTCAGTCTC
GAGGAGGCTTGATCCAGCCTGG CACTCTCCCTGCCCGTCACCCT
GGGGTCCCTGAGACTCTCCTGT TGGACAGCCGGCCTCCATCTCC
GCAGCCTCTGGGTTCACCGTCA TGCAGGTCTAGTCAAAGCCTCG
GTAGCAACTACATGAGCTGGGT TATACAGTGATGGAAACACCT CCGCCAGGCTCCAGGGAAGGG
ACTTGAATTGGTTTCAGCAGAG GCTGGAGTGGGTCTCAGTTATT GCCAGGCCAATCTCCAAGGCG
TATAGCGGTGGTAGCACATACT CCTAATTTATAAGGTTTCTAAC
ACGCAGACTCCGTGAAGGGCCG TGGGACTCTGGGGTCCCAGAC ATTCACCATCTCCAGAGACAAT
AGATTCAGCGGCAGTGGGTCA TCCAAGAACACGCTGTATCTTC GGCACTGATTTCACACTGAAA
AAATGAACAGCCTGAGAGCCG ATCAGCAGGGTGGAGGCTGAG AGGACACGGCCGTGTATTACTG
GATGTTGGGGTTTATTACTGCA TGCGAGAGAATTGTACGAAGTG TGCAACGGTACACACTGGCCG
GGAGCTACGGACTACTGGGGCC GCGTTTTCGGCCCTGGGACCAA
AGGGAACCCTGGTCACCGTCTC AGTGGATATCAAA CTCA P2B-1A10 SEQ ID NO: 174
SEQ ID NO: 175 GAGGTGCAGCTGGTGGAGTCTG GACATCCAGATGACCCAGTCTC
GAGGAGGCTTGATCCAGCCTGG CATCCTCCCTGTCTGCATCTGT
GGGGTCCCTGAGACTCTCCTGT AGGAGACAGAGTCACCATCAC GCAGCCTCTGGGTTCACCGTCA
TTGCCAGGCGAGTCAGGACAT GTAGCAACTACATGAGCTGGGT TAGCAACTATTTTAATTGGTAT
CCGCCAGGCTCCAGGGAAGGG CAGCAGAAACCAGGGAAAGCC GCTGGAGTGGGTCTCAGTTATT
CCTAAGCTCCTGATCTACGATG TATAGCGGTGGTAGCACATACT CATCCAATTTGGAAACAGGGG
ACGCAGACTCCGTGAAGGGCCG TCCCATCAAGGTTCAGTGGAA ATTCACCATCTCCAGAGACAAT
GTGGATCTGGGACAGATTTTAC TCCAAGAACACGCTGTATCTTC
TTTCACCATCAGCAGCCTGCAG AAATGAACAGCCTGAGAGCCG CCTGAAGATATTGCAACATATT
AGGACACGGCCGTTTATTACTG ACTGTCAACAGTATGATAATCT
TGCGAGAGAGGGCCCAAAGTCT CCCCATGTACACTTTTGGCCAG
ATTACAGGGACGGCTTTTGATA GGGACCAAGCTGGAGATCAAA TCTGGGGCCAAGGGACAATTGT
CACCGTCTCCTCA P2B-1D9 SEQ ID NO: 184 SEQ ID NO: 185
CAGATCACCTTGAAGGAGTCTG CAGTCTGTGCTGACTCAGCCAC
GTCCTACGCTGGTGAAACCCAC CCTCAGCGTCTGGGACCCCCGG
ACAGACCCTCACGCTGACCTGC GCAGAGGGTCACCATCTCTTGT
ACCTTCTCTGGGTTCTCACTCAG TCTGGAAGCAGCTCCAACATC
CACTAGTGGAGTGGGTGTGGGC GGAAGTAATTATGTATACTGGT
TGGATCCGTCAGCCCCCAGGAA ACCAGCAGCTCCCAGGAACGG AGGCCCTGGAGTGGCTTGCACT
CCCCCAAACTCCTCATCTATAG CATTTATTGGGATGATGATAAA TAATAATCAGCGGCCCTCAGG
TACTACAGCCCATCTCTGAAGA GGTCCCTGACCGATTCTCTGGC
GCAGGCTCACCATCACCAAGGA TCCAAGTCTGGCACCTCAGCCT
CACCTCCAAAAACCAGGTGGTC CCCTGGCCATCAGTGGGCTCCG
CTTACAATGACCAACATGGACC GTCCGAGGATGAGGCTGATTA CTGTGGACACAGCCACATATTA
TTACTGTGCAGCATGGGATGAC CTGTGCACACACTCGCATCTTA
AGCCTGAGTGGTGTGGTATTCG TACTATGGTTCGGGGAGTTATT GCGGAGGGACCAAGCTGACCG
ATGACTACTGGGGCCAGGGAAC TCCTA CCTGGTCACCGTCTCCTCA P2B-1E4 SEQ ID NO:
194 SEQ ID NO: 195 CAGATCACCTTGAAGGAGTCTG CAGTCTGCCCTGACTCAGCCTG
GTCCTACGCTGGTGAAACCCAC CCTCCGTGTCTGGGTCTCCTGG
ACAGACCCTCACGCTGACCTGC ACAGTCGATCACCATCTCCTGC
ACCTTCTCTGGGTTCTCACTCAG ACTGGAACCAGCAGTGACGTT
CACTAGTGGAGTGGGTGTGGGC GGTGGTTATAACTATGTCTCCT
TGGATCCGTCAGCCCCCAGGAA GGTACCAACAGCACCCAGGCA AGGCCCTGGAGTGGCTTGCACT
AAGCCCCCAAACTCATGATTTA CATTTATTGGGATGATGATAAG
TGATGTCAGTAAGCGGCCCTCA CGCTACAGCCCATCTCTGAAGA
GGGGTTTCTAATCGCTTCTCTG GCAGGCTCACCATCACCAAGGA GCTCCAAGTCTGGCAACACGG
CACCTCCAAAAACCAGGTGGTC CCTCCCTGACCATCTCTGGGCT
CTTACAATGACCAACATGGACC CCAGGCTGAGGACGAGGCTGA CTGTGGACACAGCCACATATTA
TTATTACTGCAGCTCATATACA CTGTGCACACCAAATAGTGGCT AGCAGCAGCGTGGTATTCGGC
ACGATTATTGACTACTGGGGCC GGAGGGACCAAGCTGACCGTC AGGGAACCCTGGTCACCGTCTC
CTA CTCA P2B-1G1 SEQ ID NO: 204 SEQ ID NO: 205
GAGGTGCAGCTGGTGGAGTCTG GAAATTGTGTTGACGCAGTCTC
GGGGAGGCTTGGTCCAGCCTGG CAGGCACCCTGTCTTTGTCTCC
GGGGTCCCTGAGACTCTCCTGT AGGGGAAAGAGCCACCCTCTC GCAGCCTCTGGATTCACCGTCA
CTGCAGGGCCAGTCAGAGTGT GTAGCAACTACATGAGCTGGGT TAGCAGCAGCTACTTAGCCTGG
CCGCCAGGCTCCAGGGAAGGG TACCAGCAGAAACCTGGCCAG GCTGGAGTGGGTCTCAGTTATT
GCTCCCAGGCTCCTCATCTATG TATAGCGGTGGTAGCACATACT GTGCATCCAGCAGGGCCACTG
ACGCAGACTCCGTGAAGGGCAG GCATCCCAGACAGGTTCAGTG ATTCACCATCTCCAGAGACAAT
GCAGTGGGTCTGGGACAGACT TCCAAGAACACGCTGTATCTTC TCACTCTCACCATCAGCAGACT
AAATGAACAGCCTGAGAGCCG GGAGCCTGAAGATTTTGCAGT AGGACACGGCTGTGTATTACTG
GTATTACTGTCAGCAGTATGGT TGCGAGAGACTACGGTGACTAC
AGCTCACCGAGGACTTTTGGCC TGGTTCGACCCCTGGGGCCAGG AGGGGACCAAGCTGGAGATCA
GAACCCTGGTCACCGTCTCCTC AA A P4A-2D9 SEQ ID NO: 214 SEQ ID NO: 215
CAGGTGCAGCTGGTGGAGTCTG GACATCCAGATGACCCAGTCTC
GGGGAGGCGTGGTCCAGCCTGG CATCCTCCCTGTCTGCATCTGT
GAGGTCCCTGAGACTCTCCTGT AGGAGACAGAGTCACCATCAC GCAGCCTCTGGATTCACCTTCA
TTGCCGGGCAAGTCAGTTCATT GTAGCTATGGCATGCACTGGGT
AGCAGCTACTTAAATTGGTATC CCGCCAGTCTCCAGGCAAGGGG AGCAGAAACCAGGGAAAGCCC
CTGGAGTGGGTGGCAGTTATAT CTAAGCTCCTGATCTATGCTAC
CAGATGATGGAAGTAATCAATA ATCCATTTTGCAAACTGGGGTC
CTATGCAGACTCCGTGAAGGGC CCATCAAGGTTCAGTGGCAGT CGATTCACCATCTCCAGAGACA
GGATCTGGGACAGATTTCACTC ATTCCAAGAACACGCTGTATCT
TCACCATCAGCAGTCTGCAACC GGAAATCAACAGCCTGAGAGTT
TGAAGATTTTGCAACTTACTAC GAGGACACGGCTGTGTATTACT
TGTCAACAGAGTTACAATACCC GTGCGAAAAGGGGCGGATATTG
TTACTTTCGGCCCTGGGACCAA TAGTACTACCAGCTGCCTCGTT AGTCGATATCAAA
AGGTGGGTCTACTTTGACTACT GGGGCCAGGGAACCCTGGTCAC CGTCTCCTCA P5A-2G7
SEQ ID NO: 224 SEQ ID NO: 225 CAGGTGCAGCTGCAGGAGTC
CAGTCTGCCCTGACTCAGCCTG GGGCCCAGGACTGGTGAAGC CCTCCGTGTCTGGGTCTCCTGG
CTTCGGAGACCCTGTCCCTCA ACAGTCGATCACCATCTCCTGC CCTGCACTGTCTCTGGTGACT
ACTGGAACCAGCAGTGACGTT CCGTCAGCAGTGGTAGTTAC GGTGGTTATAACTATGTCTCCT
TACTGGAGCTGGATCCGGCA GGTACCAACAACACCCAGGCA GCCCCCAGGGAAGGGACTGG
AAGCCCCCAAACTCATGATTTA AGTGGATTGGGTATATCTATT TGATGTCAGTAATCGGCCCTCA
ACAGTGGGAGCACCAACTAC GGGGTTTCTAATCGCTTCTCTG AACCCCTCCCTCAAGAGTCG
GCTCCAAGTCTGGCAACACGG AGTCACCATATCAGTAGACA CCTCCCTGACCATCTCTGGGCT
CGTCCAAGAACCAGTTCTCC CCAGGCTGAGGACGAGGCTGA CTGAAGCTGAGCTCTGTGAC
TTATTACTGCAGCTCATATACA CGCTGCGGACACGGCCGTGT AGCAGCAGCACTCTCGTGGTAT
ATTACTGTGCGAGAGAGCGA TCGGCGGAGGGACCAAGCTGA TGTTACTATGGTTCAGGGAG
CCGTCCTA AGCCCCCCGTTGTGTCTGGTT CGACCCCTGGGGCCAGGGAA
CCCTGGTCACCGTCTCCTCA P5A-3C8 SEQ ID NO: 234 SEQ ID NO: 235
GAGGTGCAGCTGGTGGAGTCTG GACATCCAGTTGACCCAGTCTC
GAGGAGGCTTGATCCAGCCTGG CATCCTCCCTGTCTGCATCTGT
GGGGTCCCTGAGACTCTCCTGT AGGAGACAGAGTCACCATCAC GCAGCCTCTGGGTTCACCGTCA
TTGCCGGGCCAGTCAGGGCATT GTAGCAACTACATGAGCTGGGT
AGCAGTTATTTAGCCTGGTATC CCGCCAGGCTCCAGGGAAGGG AGCAAAAACCAGGGAAAGCCC
GCTGGAATGGGTCTCATTTATTT CTAAGCTCCTGATCTATGCTGC
ATAGCGGTGGTAGTACATACTA ATCCACTTTGCAAAGTGGGGTC
CGCAGACTCCGTGAAGGGCCGA CCATCAAGGTTCAGCGGCAGT TTCACCATCTCCAGAGACAATT
GGATCTGGGACAGATTTCACTC CCAAGAACACGCTGTATCTTCA
TCACCATCAGCAGCCTGCAGCC AATGAACAGCCTGAGAGCCGA TGAAGATTTTGCAACTTATTAC
GGACACGGCCGTGTATTACTGT TGTCAACACCTTAATAGTTACC GCGAGAGATCTACAGGAACAC
CTCCGGGGTACACTTTTGGCCA GGTATGGACGTCTGGGGCCAAG GGGGACCAAGCTGGAGATCAA
GGACCACGGTCACCGTCTCCTC A A P5A-1D2 SEQ ID NO: 244 SEQ ID NO: 245
GAGGTGCAGCTGGTGGAGTCTG CAGTCTGTGCTGACGCAGCCGC
GAGGAGGCTTGATCCAGCCTGG CCTCAGTGTCTGGGGCCCCAGG
GGGGTCCCTGAGACTCTCCTGT GCAGAGGGTCACCATCTCCTGC
GCAGCCTCTGGGTTCATCGTCA ACTGGGAGCAGCTCCAACATC GTAGCAACTACATGAGCTGGGT
GGGGCAGGTTATGATGTACAC CCGCCAGGCTCCAGGGAAGGG TGGTACCAGCAACTTCCAGGA
GCTGGAGTGGGTCTCAATTATT ACAGCCCCCAAACTCCTCATCT
TATAGCGGTGGTAGCACATACT ATGGTAACAGCAATCGGCCCT ACGCAGACTCCGTGAAGGGCCG
CAGGGGTCCCTGACCGATTCTC ATTCACCATCTCCAGAGACAAT
TGGCTCCAAGTCTGGCACCTCA TCCAACAACACGCTGTATCTTC
GCCTCCCTGGCCATCACTGGGC AAATGAACAGCCTGAGAGCCG TCCAGGCTGAAGATGAGACTG
AGGACACGGCCGTATATTACTG ATTATTACTGCCAGTCCTGTGA
TGCGAGAGCCCTCCAGGTGGGA CAGCAGCCTGAGTGTTGTGGTA
GCTACTTCGGACTACTTTGACT TTCGGCGGAGGGACCAAGCTG ACTGGGGCCAGGGAACCCTGGT
ACCGTCCTA CACCGTCTCCTCA P5A-2F11 SEQ ID NO: 254 SEQ ID NO: 255
CAGGTGCAGCTGGTGCAGTCTG GACATCGTGATGACCCAGTCTC GGGCTGAGGTGAAGAAGCCTG
CAGACTCCCTGGCTGTGTCTCT GGGCCTCAGTGAAGGTCTCCTG GGGCGAGAGGGCCACCATCAA
CAAGGCTTCTGGATACACCTTC CTGCAAGTCCAGCCAGAGTGTT
ACCAGTTATGATATCAACTGGG TTATACAGCTCCAACAATAAG TGCGACAGGCCACTGGACAAGG
AACTACTTAGCTTGGTACCAGC GCTTGAGTGGATGGGATGGATG AGAAACCAGGACAGCCTCCTA
AACCCTAACAGTGGTAACACAG AGCTGCTCATTTACTGGGCATC
GCTATGCACAGAAGTTCCAGGG TACCCGGGAATCCGGGGTCCCT
CAGAGTCACCATGACCAGGAAC GACCGATTCAGTGGCAGCGGG ACCTCCATAAGCACAGCCTACA
TCTGGGACAGATTTCACTCTCA TGGAGCTGAGCAGCCTGAGATC CCATCAGCAGCCTGCAGGCTG
TGAGGACACGGCCGTGTATTAC AAGATGTGGCAGTTTATTACTG
TGTGCGAGATATATTGTAGTAG TCAGCAATATTATAGTACTCCT
TACCAGCTGCAAAAGGGTTCGA CTCACTTTCGGCGGAGGGACC CCCCTGGGGCCAGGGAACCCTG
AAGGTGGAGATCAAA GTCACCGTCTCCTCA P5A-2E1 SEQ ID NO: 264 SEQ ID NO:
265 GAGGTGCAGCTGGTGCAGTCTG TCCTATGTGCTGACTCAGCCAC
GAGCAGAGGTGAAAAAGCCCG CCTCAGTGTCAGTGGCCCCAGG GGGAGTCTCTGAAGATCTCCTG
AAAGACGGCCAGGATTACCTG TAAGGGTTCTGGATACAGCTTT TGGGGGAAACAACATTGGAAG
ACCAGCTACTGGATCGGCTGGG TAAAAGTGTGCACTGGTACCA TGCGCCAGATGCCCGGGAAAGG
GCAGAAGCCAGGCCAGGCCCC CCTGGAGTGGATGGGGATCATC TGTGCTGGTCATCTATTATGAT
TATCCTGGTGACTCTGATACCA AGCGACCGGCCCTCAGGGATC GATACAGCCCGTCCTTCCAAGG
CCTGAGCGATTCTCTGGCTCCA CCAGGTCACCATCTCAGCCGAC ACTCTGGGAACACGGCCACCC
AAGTCCATCAGCACCGCCTACC TGACCATCAGCAGGGTCGAAG TGCAGTGGAGCAGCCTGAAGGC
CCGGGGATGAGGCCGACTATT CTCGGACACCGCCATGTATTAC ACTGTCAGGTGTGGGATAGTA
TGTGCCCAGACGTCAGTGACTC GTAGTGATCATGTGGTATTCGG
GCAACTGGTTCGACCCCTGGGG CGGAGGGACCAAGCTGACCGT CCAGGGAACCCTGGTCACCGTC
CCTA TCCTCA P5A-1C8 SEQ ID NO: 274 SEQ ID NO: 275
CAGGTGCAGCTGGTGCAGTCTG GACATCCAGATGACCCAGTCTC GGGCTGAGGTGAAGAAGCCTG
CATCCTCCCTGTCTGCATCTGT GGGCCTCAGTGAAGGTTTCCTG AGGAGACAGAGTCACCATCAC
CAAGGCATCTGGATACACCTTC TTGCCAGGCGAGTCAGGACAT ACCAGCTACTATATGCACTGGG
TAGCAACTATTTAAATTGGTAT TGCGACAGGCCCCTGGACAAGG CAGCAGAAACCAGGGAAAGCC
GCTTGAGTGGATGGGAATAATC CCTAAGCTCCTGATCTACGATG
AACCCTAGTGGTGGTAGCACAA CATCCAATTTGGAAACAGGGG GCTACGCACAGAAGTTCCAGGG
TCCCATCAAGGTTCAGTGGAA CAGAGTCACCATGACCAGGGAC GTGGATCTGGGACAGATTTTAC
ACGTCCACGAGCACAGTCTACA TTTCACCATCAGCAGCCTGCAG
TGGAGCTGAGCAGCCTGAGATC CCTGAAGATATTGCAACATATT
TGAGGACACGGCCGTGTATTAC ACTGTCAACAGTATGATAATCT
TGTGCGAGGTCGGCCCGGGATT CCCCTCTATCACCTTCGGCCAA
ACTATGATAGTAGTGGTTATTA GGGACACGACTGGAGATTAAA CTACCGCGCTGAATACTTCCAG
CACTGGGGCCAGGGCACCCTGG TCACCGTCTCCTCA P1A-1C10 SEQ ID NO: 284 SEQ
ID NO: 285 CAGGTGCAGCTGGTGCAGTCTG GACATCCAGATGACCCAGTCTC
GGGCTGAGGTGAAGAACCCGG CTTCCACCCTGTCTGCATCTGT GGTCCTCGGTGAAGGTCTCCTG
AGGAGACAGAGTCACCATCAC TAAGGCTGGTGGAGGCACCTCC TTGCCGGGCCAGTCAGAGTTCT
AGTTTCTATGATATCAACTGGG AGGGCCTGGTTGGCCTGGTATC
TGCGACAGGCCCCTGGACAAGG AGCAGAAACCAGGGAAAGCCC GCTTGAGTGGATAGGAAAAATC
CTAAACTCCTGATCTCTAAGGC ATCCCTAGGCTTGATATAGCAG
GTCTAGTTTAGAAAGTGGGGTC ACTACGCACAGAAGTCCCAGGG CCATCAAGGTTCAGCGGCAGT
CAGAGTCACGATTACCGCGGAC GGATATGGGACAGAATTCACT AAATCCACGAGTACAGTATACT
CTCACCATCAGCAGCCTGCAGC TGGAATTGAGCAGCCTGAAGTC
CTGATGATTCTGCAACTTATTA AGACGACACGGCCGTGTATTTC CTGCCACCAGTATAACAGTAG
TGTGCGAGAGGTCGGCCGGGTT CCCATTCACTTTCGGCCCTGGG
CGGAGTGGGCGTATGGCCCATT ACCAAAGTGCAGATCAAA TGACCTCTGGGGCCAGGGAACC
CTGGTCACCGTCTCCTCA P4A-1H6 SEQ ID NO: 294 SEQ ID NO: 295
CAGGTGCAGCTGGTGGAGTCTG GACATCCAGATGACCCAGTCTC
GGGGAGGCGTGGTCCAGCCTGG CATCCTCCCTGTCTGCATCTGT
GAGGTCCCTGAGACTCTCCTGT AGGAGACAGAGTCACCATCAC GCAGCCTCTGGATTCACCTTCA
TTGCCGGGCAAGTCAGAGCAT GTAGCTATGGCATGCACTGGGT TAGCAGCTATTTACATTGGTAT
CCGCCAGTCTCCAGGCAAGGGG CAGCAAAAACCAGGGAAAGCC CTGGAGTGGGTGGCAGTTATAT
CCTAACCTCCTGATCTATGCTG CAGATGATGGAAGTAATCAATA
CATCCAGTTTGCAAAGTGGGGT CTATGCAGACTCCGTGAAGGGC
CCCATCAAGGTTCAGTGGCAGT CGATTCACCATCTCCAGAGACA
GGATCTGGGACAGATTTCACTC ATTCCAAGAACACGCTGTATCT
TCACCATCAGCAGTCTGCAACC GCAAATGAACAGCCTGAGAGTT
TGAAGACTTTGCAACTTACTAC GAGGACACGGCTGTGTATTACT
TGTCAACAGAGTTACAATACCC GTGCGAAAAGGGGCGGATATTG
CTACTTTCGGCCCTGGGACCAA TAGTACTACCAGCTGCCTCCTT AGTGGATATCAAA
AGGTGGGTCTACTTTGACTTCT GGGGCCAGGGAACCCTGGCCAC CGTCTCCTCA P4B-1F4
SEQ ID NO: 304 SEQ ID NO: 305 CAGGTGCAGCTGGTGGAGTCTG
GATGTTGTGATGACTCAGTCTC GGGGAGGCGTGGTCCAGCCTGG
CACTCTCCCTGCCCGTCACCCT GAGGTCCCTGAGACTCTCCTGT
TGGACAGCCGGCCTCCATCTCC GCAGCCTCTGGATTCACCTTCA
TGCAGGTCTAGTCAAAGCCTCG GTAGCTATGGCATGCACTGGGT TATACAGTGATGGAAACACCT
CCGCCAGGCTCCAGGCAAGGGG ACTTGAATTGGTTTCAGCAGAG
CTGGAGTGGGTGGCAGTTATAT GCCAGGCCAATCTCCAAGGCG CATATGATGGAAGTAATAAATA
CCTAATTTATAAGGTTTCTAAC CTATGCAGACTCCGTGAAGGGC CGGGACTCTGGGGTCCCAGAC
CGATTCACCATCTCCAGAGACA AGATTCAGCGGCAGTGGGTCA ATTCCAAGAACACGCTGTATCT
GGCACTGATTTCACACTGAAA GCAAATCAACAGCCTGAGAGCT ATCAGCAGGGTGGAGGCTGAG
GAGGACACGGCTGTGTATTACT GATGTTGGGGTTTATTACTGCA
GTGCGAAAGGGCCTCGGTATAG TGCAAGCTACACACTGGCCCCT
CAGCAGCTGGTACATAAGCCTT GTACACTTTTGGCCAGGGGACC
TACTACTACTACGGTATGGACG AAGCTGGAGATCAAA TCTGGGGCCAAGGGACCACGGT
CACCGTCTCCTCA P5A-1B6 SEQ ID NO: 314 SEQ ID NO: 315
CAGGTGCAGCTGGTGGAGTCTG GACATCCAGATGACCCAGTCTC
GGGGAGGCGTGGTCCAGCCTGG CATCCTCCCTGTCTGCATCTGT
GAGGTCCCTGAGACTCTCCTGT AGGAGACAGAGTCACCATCAC GCAGCCTCTGGATTCACCTTCA
TTGCCAGGCGAGTCAGGACAT GTAGCTATGCTATGCACTGGGT TAGCAACTATTTAAATTGGTAT
CCGCCAGGCTCCAGGCAAGGGG CAGCAGAAACCAGGGAAAGCC CTGGAGTGGGTGGCAGTTATAT
CCTAAGCTCCTGATCTACGATG CATATGATGGAAGTAATAAATA CATCCAATTTGGAAACAGGGG
CTACGCAGACTCCGTGAAGGGC TCCCATCAAGGTTCAGTGGAA CGATTCACCATCTCCAGAGACA
GTGGATCTGGGACAGATTTTAC ATTCCAAGAACACGCTGTATCT
TTTCACCATCAGCAGCCTGCAG GCAAATGAACAGCCTGAGAGCT
CCTGAAGATATTGCAACATATT GAGGACACGGCTGTGTATTACT
ACTGTCAACAGTATGATAATCT GTGCGAGAGATGGACAGGCTAT
CCCGTACACTTTTGGCCAGGGG TACTATGGTTCAGGGAGTTATC ACCAAGCTGGAGATCAAA
GGCCCACCCTTTGACTACTGGG GCCAGGGAACCCTGGTCACCGT CTCCTCA P5A-1B8 SEQ
ID NO: 324 SEQ ID NO: 325 GAGGTGCAGCTGGTGGAGTCTG
GACATCCAGTTGACCCAGTCTC GAGGAGGCTTGATCCAGCCTGG
CATCCTTCCTGTCTGCATCTGT GGGGTCCCTGAGACTCTCCTGT AGGAGACAGAGTCACCATCAC
GCAGCCTCTGGGTTCACCGTCA TTGCCGGGCCAGTCAGGGCATT
GTAGCAACTACATGAGCTGGGT AGCAGTTATTTAGCCTGGTATC CCGCCAGGCTCCAGGGAAGGG
AGCAAAAACCAGGGAAAGCCC GCTGGAGTGGGTCTCAGTTATT CTAAGCTCCTGATCTATGCTGC
TATCCCGGTGGTAGCACATTCT ATCCACTTTGCAAAGTGGGGTC
ACGCAGACTCCGTGAAGGGCCG CCATCAAGGTTCAGCGGCAGT ATTCACCATCTCCAGAGACAAT
GGATCTGGGACAGAATTCACT TCCAAGAACACCCTGTATCTTC CTCACAATCAGCAGCCTGCAG
AAATGAACAGCCTGAGAGCCG CCTGAAGATTTTGCAACTTATT AGGACACGGCCGTGTATTACTG
ACTGTCAACAGCTTAATAGTTA TGCGAGAGAGACCCTAGCCTTT
CCCTCCAGCTTTCGGCGGAGGG GACTACTGGGGCCAGGGAACCC ACCAAGGTGGAGATCAAA
TGGTCACCGTCTCCTCA PSA-1B9 SEQ ID NO: 334 SEQ ID NO: 335
CAGGTGCAGCTGCAGGAGTCGG GACATCGTGATGACCCAGTCTC
GCCCAGGACTGGTGAAGCCTTC CAGACTCCCTGGCTGTGTCTCT
GGAGACCCTGTCCCTCACCTGC GGGCGAGAGGGCCACCATCAA ACTGTCTCTGGTGGCTCCATCA
CTGCAAGTCCAGCCAGAGTGTT GTAGTTACTACTGGAGCTGGAT TTATACAGCTCCAACAATAAG
CCGGCAGCCCCCAGGGAAGGG AACTACTTAGCTTGGTACCAGC ACTGGAGTGGATTGGGTATATC
AGAAACCAGGACAGCCTCCTA TCTTACAGTGGGAGCACCAACT AGCTGCTCATTTACTGGGCATC
ACAACCCCTCCCTCAAGAGTCG TACCCGGGAATCCGGGGTCCCT
AGTCACCATATCACTAGACACG GACCGATTCAGTGGCAGCGGG TCCAAGAACCAGTTCTCCCTGA
TCTGGGACAGATTTCACTCTCA AGCTGAGCTCTGTGACCGCTGC CCATCAGCAGCCTGCAGGCTG
GGACACGGCCGTGTATTACTGT AAGATGTGGCAGTTTATTACTG
GCGAGCAACGGCCAGTATTACG TCAGCAATATTATAGTACTCCG
ATATTTTGACTGGTCAACCTCCT CTCACTTTCGGCGGAGGGACC
GACTACTGGTACTTCGATCTCT AAGGTGGAGATCAAA GGGGCCGTGGCACCCTGGTCAC
TGTCTCCTCA P5A-1D1 SEQ ID NO: 344 SEQ ID NO: 345
GAGGTGCAGCTGGTGGAGTCTG GACATCCAGTTGACCCAGTCTC
GAGGAGGCTTGATCCAGCCTGG CATCCTTCCTGTCTGCATCTGT
GGGGTCCCTGAGACTCTCCTGT AGGAGACAGAGTCACCATCAC GCAGCCTCTGGGCTCACCGTCA
TTGCCGGGCCAGTCAGGGCATT GTAGCAACTACATGAGCTGGGT
AGCAGTTATTTAGCCTGGTATC CCGCCAGGCTCCAGGGAAGGG AGCAAAAACCAGGGAAAGCCC
GCTGGAGTGGGTCTCAGTTATT CTAAGCTCCTGATCTATGCTGC
TATAGCGGTGGTAGCACATACT ATCCACTTTGCAAAGTGGGGTC
ACGCAGACTCCGTGAAGGGCCG CCATCAAGGTTCAGCGGCAGT ATTCACCATCTCCAGAGACAAT
GGATCTGGGACAGATTTCACTC TCCAAGAACACGCTGTATCTTC
TCACCATCAGCAGCCTGCAGCC AAATGAACAGCCTGAGAGCCG TGAAGATTTTGCAACTTATTAC
AGGACACGGCCGTGTATTACTG TGTCAACAGCTTAATAGTTACC
TGCGAGAGATTTGTACTACTAC CTACCTTCGGCCAAGGGACAC GGTATGGACGTCTGGGGCCAAG
GAC TGGAGATTAAA GGACCACGGTCACCGTCTCCAC A PSA-1D10 SEQ ID NO: 354
SEQ ID NO: 355 CAGGTGCAGCTGGTGGAGTCTG CAGTCTGCCCTGACTCAGCCTG
GGGGAGGCTTGGTCAAGCCTGG CCTCCGTGTCTGGGTCTCCTGG
AGGGTCCCTGAGACTCTCCTGT ACAGTCGATCACCATCTCCTGC
GCAGCCTCTCAATTCACCTTCA ACTGGAACCAGCAGTGACGTT GTGACTACTCCATGACCTGGAT
GGTGGTTATAACTATGTCTCCT CCGCCAGGCTCCAGGGAAGGG GGTACCAACAACACCCAGGCA
GCTGGAGTGGGTTTCATACATT AAGCCCCCAAACTCATGATTTA
AGTCAAAGTGGTAGTACCATAT TGATGTCAGTAATCGGCCCTCA
ACTACGCAGACTCTGTGAAGGG GGGGTTTCTAATCGCTTCTCTG
CCGATTCACCATCTCCAGGGAC CCTCCAAGTCTGGCAACACGG AACGCCAAGAACTCACTGTATC
CCTCCCTGACCATCTCTGGGCT TGCAAATGAACAGCCTGAGAGC CCAGGCTGAGGACGAGGCTGA
CGAGGACACGGCCGTGTATTAC TTATTACTGCAGCTCATTTACA
TGTGCGAGAGGTGTCAGCCCAT AGCAGCACCACTGTCGTGGTAT
CCTACGTTTGGGGGAGTTATCG TCGGCGGAGGGACCAAGCTGA
TTCCTTGTACCACTTTGACTACT CCGTCCTA GGGGCCAGGGAACCCTGGTCAC CGTCTCCTCA
P5A-2D11 SEQ ID NO: 364 SEQ ID NO: 365 GAGGTGCAGCTGGTGCAGTCTG
CAGTCTGTGCTGACTCAGCCAC GAGCAGAGGTGAAAAAGCCCG CCTCAGCGTCTGGGACCCCCGG
GGGAGTCTCTGAAGATCTCCTG GCAGAGGGTCACCATCTCTTGT
TAAGGGTTCTGGATACAGCTTT TCTGGAAGCAGCTCCAACATC ACCAGCTACTGGATCGGCTGGG
GGAAGTAATACTGTAAACTGG TGCGCCAGATGCCCGGGAAAGG TACCAGCAGCTCCCAGGAACG
CCTGGAGTGGATGGGGATCATC GCCCCCAAACTCCTCATCTATA
TATCCTGGTGACTCTGATACCA GTAATAATCAGCGGCCCTCAG GATACAGCCCGTCCTTCCAAGG
GGGTCCCTGACCGATTCTCTGG CCAGGTCACCATCTCAGCCGAC
CTCCAAGTCTGGCACCTCAGCC AAGTCCATCAGCACCGCCTACC
TCCCTGGCCATCAGTGGGCTCC TGCAGTGGAGCAGCCTGAAGGC AGTCTGAGGATGAGGCTGATT
CTCGGACACCGCCATGTATTAC ATTACTGTGCAGCATGGGATG TGTGCGAGACGGGATTCGACCT
ACAGCCTGAATGGTGTGGTATT ACGGTGGTAACACTGACTACTG CGGCGGAGGGACCAAGCTGAC
GGGCCAGGGAACCCTGGTCACC CGTCCTA GTCTCCTCA P5A-2G9 SEQ ID NO: 374 SEQ
ID NO: 375 CAGGTGCAGCTGGTGGAGTCTG CAGCCTGTGCTGACTCAGCCAC
GGGGAGGCGTGGTCCAGCCTGG CTTCCTCCTCCGCATCTCCTGG
GAGGTCCCTGAGACTCTCCTGT AGAATCCGCCAGACTCACCTG GCAGCGTCTGGATTCACCTTCA
CACCTTGCCCAGTGACATCAAT GTAGCTATGGCATGCACTGGGT
GTTAGTAGCTACAACATATACT CCGCCAGGCTCCAGGCAAGGGG GGTACCAGCAGAAGCCAGGGA
CTGGAGTGGGTGGCAGTTATAT GCCCTCCCAGGTATCTCCTGTA
GGTATGATGGAAGTAATAAATA CTACTACTCAGACTCAGATAAG
CTATGCAGACTCCGTGAAGGGC GGCCAGGGCTCTGGAGTCCCC CGATTCACCATCTCCAGAGACA
AGCCGCTTCTCTGGATCCAAAG ATTCCAAGAACACGCTGTATCT ATGCTTCAGCCAATACAGGGA
GCAAATGAACAGCCTGAGAGCC TTTTACTCATCTCCGGGCTCCA
GAGGACACGGCTGTGTATTACT GTCTGAGGATGAGGCTGACTA GTGCGAGATGGTTCCACACGGG
TTACTGTATGATTTGGCCAAGC GGGGTACTTTGACTACTGGGGC
AATGCTCTTTATGTCTTCGGAA CAGGGAACCCTGGTCACCGTCT
CTGGGACCAAGGTCACCGTCCT CCTCA A PSA-2H3 SEQ ID NO: 384 SEQ ID NO:
385 GAGGTGCAGCTGGTGCAGTCTG CAGTCTGTGCTGACTCAGCCAC
GAGCAGAGGTGAAAAAGCCCG CCTCAGCGTCTGGGACCCCCGG GGGAGTCTCTGAAGATCTCCTG
GCAGAGGGTCACCATCTCTTGT TAAGGGTTCTGGATACAGCTTT TCTGGAAGCAGCTCCAACATC
ACCAGCTACTGGATCGGCTGGG GGAAGTAATACTGTAAACTGG TGCGCCAGATGCCCGGGAAAGG
TACCAGCAGCTCCCAGGAACG CCTGGAGTGGATGGGGATCATC GCCCCCAAACTCCTCATCTATA
TATCCTGGTGACTCTGATACCA GTAATAATCAGCGGCCCTCAG GATACAGCCCGTCCTTCCAAGG
GGGTCCCTGACCGATTCTCTGG CCAGGTCACCATCTCAGCCGAG
CTCCAAGTCTGGCACCTCAGCC AAGTCCATCAGCACCGCCTACC
TCCCTGGCCATCAGTGGGCTCC TGCAGTGGAGCAGCCTGAAGGC AGTCTGAGGATGAGGCTGATT
CTCGGACACCGCCATGTATTAC ATTACTGTGCAGCATGGGATG TGTGCGAGACGGGATTCGACCT
ACAGCCTGAATGGTGTGGTATT ACGGTGGTAACACTGACTACTG CGGCGGAGGGACCAAGCTGAC
GGGCCAGGGAACCCTGGTCACC CGTCCTA GTCTCCTCA P5A-3A1 SEQ ID NO: 394 SEQ
ID NO: 395 GAGGTGCAGCTGGTGGAGTCTG GAAATTGTGTTGACGCAGTCTC
GAGGAGGCTTGATCCAGCCTGG CAGGCACCCTGTCTTTGTCTCC
GGGGTCCCTGAGACTCTCCTGT AGGGGAAAGAGCCACCCTCTC GCAGCCTCTGGGTTCACCGTCA
CTGCAGGGCCAGTCAGAGTGT GTAGCAACTACATGAGCTGGGT TAGCAGCAGCTACTTAGCCTGG
CCGCCAGGCTCCAGGGAAGGG TACCAGCAGAAACCTGGCCAG GCTGGAGTGGGTCTCAGTTATT
GCTCCCAGGCTCCTCATCTATG TATAGCGGTGGTAGCACATACT GTGCATCCAGCAGGGCCACTG
ACGCAGACTCCGTGAAGGGCCG GCATCCCAGACAGGTTCAGTG ATTCACCATCTCCAGAGACAAT
GCAGTGGGTCTGGGACAGACT TCCAAGAACACGCTGTATCTTC TCACTCTCACCATCAGCAGACT
AAATGAACAGCCTGAGAGCCG GGAGCCTGAAGATTTTGCAGT AGGACACGGCCGTGTATTACTG
GTATTACTGTCAGCAGTATGGT TGCGAGAGACTACGGTGACTTT
AGCTCACCTCGCACTTTTGGCC TACTTTGACTACTGGGGCCAGG AGGGGACCAAGCTGGAGATCA
GAACCCTGGTCACCGTCTCCTC AA A PSA-3A6 SEQ ID NO: 404 SEQ ID NO: 405
GAAGTGCAGCTGGTGGAGTCTG CAGTCTGCCCTGACTCAGCCTG
GGGGAGGCTTGGTACAGCCTGG CCTCCGTGTCTGGGTCTCCTGG
CAGGTCCCTGAGACTCTCCTGT ACAGTCGATCACCATCTCCTGC
GCAGCCTCTGGATTCACCTTTG ACTGGAACCAGCAGTGACGTT ATGATTATGCCATGCACTGGGT
GGTGGTTATAACTATGTCTCCT CCGGCAAGCTCCAGGGAAGGG GGTACCAACAACACCCAGGCA
CCTGGAGTGGGTCTCAGGTATT AAGCCCCCAAACTCATGATTTA
AGTTGGAATAGTGGTACCATAG TGATGTCAGTAATCGGCCCTCA
GCTATGCGGACTCTGTGAAGGG GGGGTTTCTAATCGCTTCTCTG
CCGATTCATCATCTCCAGAGAC GCTCCAAGTCTGGCAACACGG AACGCCAAGAACTCCCTGTATC
CCTCCCTGACCATCTCTGGGCT TGCAAATGAACAGTCTGAGAGC CCAGGCTGAGGACGAGGCTGA
TGAGGACACGGCCTTGTATTAC TTATTACTGCAGCTCATATACA
TGTGCAGGGGGTGGTACTATGG AGCAGCAGCACTGTGGTATTC TTCGGGGAGTTATTGCCGGAGG
GGCGGAGGGACCAAGCTGACC GGGAACTCATCCGGTGGATGAC GTCCTA
TACTACGGTATGGACGTCTGGG GCCAAGGGACCACGGTCACCGT CTCCTCA P5A-3B4 SEQ
ID NO: 414 SEQ ID NO: 415 GAGGTGCAGCTGGTGCAGTCTG
CAGTCTGTGCTGACTCAGCCAC GAGCAGAGGTGAAAGAGCCCG CCTCAGCGTCTGGGACCCCCGG
GGGAGTCTCTGAAGATCTCCTG GCAGAGGGTCACCATCTCTTGT
TAAGGGTTCTGGATACAGCTTT TCTGGAAGCAGCTCCAACATC ACCAGCTACTGGATCGGCTGGG
GGAAGTAATACTGTAAACTGG TGCGCCAGATGCCCGGGAAAGG TACCAGCAGCTCCCAGGAACG
CCTGGAGTGGATGGGGATCATC GCCCCCAAACTCCTCATCTATA
TATCCTGGTGACTCTGATACCA GTAATAATCAGCGGCCCTCAG GATACAGCCCGTCCTTCCAAGG
GGGTCCCTGACCGATTCTCTGG CCAGGTCACCATCTCAGCCGAC
CTCCAAGTCTGGCACCTCAGCC AAGTCCATCAGCACCGCCTACC
TCCCTGGCCATCAGTGGGCTCC TGCAGTGGAGCAGCCTGAAGGC AGTCTGAGGATGAGGCTGATT
CTCGGACACCGCCATGTATTAC ATTACTGTGCAGCATGGGATG TGTGCGAGACGGGATTCGACCT
ACAGCCTGAATGGTGTGGTATT ACGGTGGTAACACTGACTACTG CGGCGGAGGGACCAAGCTGAC
GGGCCAGGGAACCCTGGTCACC CGTCCTA GTCTCCTCA PSA-3C12 SEQ ID NO: 424
SEQ ID NO: 425 CAGATCACCTTGAAGGAGTCTG GACATCGTGATGACCCAGTCTC
GTCCTACGCTGGTGAAACCCAC CAGACTCCCTGGCTGTGTCTCT
ACAGACCCTCACGCTGACCTGC GGGCGAGAGGGCCACCATCAA
ACCTTCTCTGGGTTCTCACTCAG CTGCAAGTCCAGCCAGAGTGTT
CACTAGTGGAGTGGGTGTGGGC TTATACAGCTCCAACAATAAG TGGATCCGTCAGCCCCCAGGAA
AACTACTTAGCTTGGTACCAGC AGGCCCTGGAGTGGCTTGCACT AGAAACCAGGACAGCCTCCTA
CATTTATTGGGATGATGATAAG AGCTGCTCATTTACTGGGCATC
CGCTACAGCCCATCTCTGAAGA TACCCGGGAATCCGGGGTCCCT
GCAGGCTCACCATCACCAAGGA GACCGATTCAGTGGCAGCGGG CACCTCCAAAAACCAGGTGGTC
TCTGGGACAGATTTCACTCTCA CTTACAATGACCAACATGGACC CCATCAGCAGCCTGCAGGCTG
CTGTGGACACAGCCACATATTA AAGATGTGGCAGTTTATTACTG
CTGTGCACACAGTTTGTTTCTCA TCAGCAATATTATAGTACTCCT
CGGTAGGGTATAGCAGCAGCTG CACACTTTTGGCCAGGGGACC GTCCCCTTTTGACTACTGGGGC
AAGCTGGAGATCAAA CAGGGAACCCTGGTCACCGTCT CCTCA P22A-1D1 SEQ ID NO:
434 SEQ ID NO: 435 GAGGTGCAGCTGGTGGAGTCTG GACATCCAGTTGACCCAGTCTC
GAGGAGGCTTGATCCAGCCTGG CATCCTTCCTGTCTGCATCTGT
GGGGTCCCTGAGACTCTCCTGT AGGAGACAGAGTCACCATCAC GCAGCCTCTGGGTTCACCGTCA
TTGCCGGGCCAGTCAGGGCATT GTAGCAACTACATGAGCTGGGT
AGCAGTTATTTAGCCTGGTATC CCGCCAGGCTCCAGGGAAGGG AGCAAAAACCAGGGAAAGCCC
GCTGGAGTGGGTCTCAGTTATT CTAAGCTCCTGATCTATGCTGC
TATAGCGGTGGTAGCACATACT ATCCACTTTGCAAAGTGGGGTC
ACGCAGACTCCGTGAAGGGCCG CCATCAAGGTTTAGCGGCAGT ATTCACCATCTCCAGAGACAAT
GGATCTGGGACAGAATTCACT TCCAAGAACACGCTGTATCTTC CTCACAATCAGCAGCCTGCAG
AAATGAACAGCCTGAGAGCCG CCTGAAGATTTTGCAACTTATT AGGACACGGCCGTGTATTACTG
ACTGTCTACACCTTAATAGTTA TGCGAGAGATCGAGACTACTAC CAGGACGTTCGGCCTAGGGAC
GGTATGGACGTCTGGGGCCAAG CAAGGTGGAAATCAAA GGACCACGGTCACCGTCTCCTC
A
[0355] In certain embodiments, the antibodies or the
antigen-binding fragments thereof provided herein further comprise
an immunoglobulin (Ig) constant region, which optionally further
comprises a heavy chain and/or a light chain constant region. In
certain embodiments, the heavy chain constant region comprises CH1,
hinge, and/or CH2-CH3 regions (or optionally CH2-CH3-CH4 regions).
In certain embodiments, the antibodies or the antigen-binding
fragments thereof provided herein comprises heavy chain constant
regions of human IgG1, IgG2, IgG3, IgG4, IgA1, IgA2 or IgM. In
certain embodiments, the light chain constant region comprises
C.kappa. or C.lamda.. The constant region of the antibodies or the
antigen-binding fragments thereof provided herein may be identical
to the wild-type constant region sequence or be different in one or
more mutations.
[0356] In certain embodiments, the heavy chain constant region
comprises an Fc region. Fc region is known to mediate effector
functions such as antibody-dependent cellular cytotoxicity (ADCC),
Antibody-dependent cellular phagocytosis (ADCP) and
complement-dependent cytotoxicity (CDC) of the antibody. Fc regions
of different Ig isotypes have different abilities to induce
effector functions. For example, Fc regions of IgG1 and IgG3 have
been recognized to induce both ADCC and CDC more effectively than
those of IgG2 and IgG4. In certain embodiments, the antibodies and
antigen-binding fragments thereof provided herein comprises an Fc
region of IgG1, or IgG3 isotype, which could induce ADCC or CDC.
Alternatively, the antibodies and antigen-binding fragments thereof
provided herein comprise a constant region of IgG4 or IgG2 isotype,
which has reduced or depleted effector function. In certain
embodiments, the anti-SARS-COV-2 antibodies or antigen-binding
fragments thereof comprises a wild type human IgG1 Fc region
comprising the sequence of SEQ ID NO: 115 or other wild type human
IgG1 alleles.
[0357] Table 4 shows the amino acid sequences for the heavy chain
and light chain constant regions of the monoclonal antibodies:
P2A-1A8, P2A-1A9, P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4, P2B-2G11,
P2C-1A3, P2C-1C8, P2C-1C10, P2C-1D5, P2C-1F11, P2B-1G5, P2B- 1A1,
P2C-1D7, P2B-1A10, P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7,
P5A-3C8, P5A-1D2, P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6,
P4B-1F4, P5A-1B6, P5A- 1B8, P5A-1B9, P5A-1D1, P5A-1D10, P5A-2D11,
P5A-2G9, P5A-2H3, P5A-3A1, P5A-3A6, P5A-3B4, P5A-3C12, and P22A-1D1
wherein the antibodies P2A-1A8, P2A-1A9, P2B-2F6, P2B-2G4,
P2B-2G11, P2C-1D5, P2B-1G5, P2B-1A1, P2B-1D9, P2B-1E4, P5A-2G7,
P5A-1D2, P5A-2E1, P5A-1D10, P5A-2D11, P5A-2G9, P5A-2H3, P5A-3A6,
and P5A-3B4 have lambda light chains (with a lambda light chain
constant region sequence of SEQ ID NO: 116), the antibodies
P2A-1A10, P2A-1B3, P2C-1A3, P2C-1C8, P2C-1C10, P2C-1F11, P2C-1D7,
P2B-1A10, P2B-1G1, P4A-2D9, P5A-3C8, P5A-2F11, P5A-1C8, P1A-1C10,
P4A-1H6, P4B- 1F4, P5A-1B6, P5A-1B8, P5A-1B9, P5A-1D1, P5A-3A1,
P5A-3C12, and P22A-1D1 have kappa light chains (with a kappa light
chain constant region sequence of SEQ ID NO: 117), and all 42
antibodies have the same heavy chain constant region (SEQ ID NO:
115).
TABLE-US-00005 TABLE 4 Amino acid and nucleic acid sequences of
constant regions HC Amino SEQ ID NO: 115 (Heavy acid
ASTKGPSVFPLAPSSKSTSGGTAALGCLVK Chain DYFPEPVTVSWNSGALTSGVHTFPAVLQSS
constant GLYSLSSVVTVPSSSLGTQTYICNVNHKPS region)
NTKVDKKVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVS
HEDPEVKFNWYVDGVEVHNAKTKPREEQYN STYRVVSVLTVLHQDWLNGKEYKCKVSNKA
LPAPIEKTISKAKGQPREPQVYTLPPSRDE LTKNQVSLTCLVKGFYPSDIAVEWESNGQP
ENNYKTTPPVLDSDGSFFLYSKLTVDKSRW QQGNVFSCSVMHEALHNHYTQKSLSLSPGK
Nucleic SEQ ID NO: 118 Acid GCGTCGACCAAGGGCCCATCGGTCTTCCCC
CTGGCACCCTCCTCCAAGAGCACCTCTGGG GGCACAGCGGCCCTGGGCTGCCTGGTCAAG
GACTACTTCCCCGAACCCGTGACGGTGTCG TGGAACTCAGGCGCCCTGACCAGCGGCGTG
CACACCTTCCCGGCTGTCCTACAGTCCTCA GGACTCTACTCCCTCAGCAGCGTGGTGACC
GTGCCCTCCAGCAGCTTGGGCACCCAGACC TACATCTGCAACGTGAATCACAAGCCCAGC
AACACCAAGGTGGACAAGAAAGTTGAGCCC AAATCTTGTGACAAAACTCACACATGCCCA
CCGTGCCCAGCACCTGAACTCCTGGGGGGA CCGTCAGTCTTCCTCTTCCCCCCAAAACCC
AAGGACACCCTCATGATCTCCCGGACCCCT GAGGTCACATGCGTGGTGGTGGACGTGAGC
CACGAAGACCCTGAGGTCAAGTTCAACTGG TACGTGGACGGCGTGGAGGTGCATAATGCC
AAGACAAAGCCGCGGGAGGAGCAGTACAAC AGCACGTACCGTGTGGTCAGCGTCCTCACC
GTCCTGCACCAGGACTGGCTGAATGGCAAG GAGTACAAGTGCAAGGTCTCCAACAAAGCC
CTCCCAGCCCCCATCGAGAAAACCATCTCC AAAGCCAAAGGGCAGCCCCGAGAACCACAG
GTGTACACCCTGCCCCCATCCCGGGATGAG CTGACCAAGAACCAGGTCAGCCTGACCTGC
CTGGTCAAAGGCTTCTATCCCAGCGACATC GCCGTGGAGTGGGAGAGCAATGGGCAGCCG
GAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTAC
AGCAAGCTCACCGTGGACAAGAGCAGGTGG CAGCAGGGGAACGTCTTCTCATGCTCCGTG
ATGCATGAGGCTCTGCACAACCACTACACG CAGAAGAGCCTCTCCCTGTCTCCGGGTAAA LC
Amino SEQ ID NO: 116 (lambda acid GQPKAAPSVTLFPPSSEELQANKATLVCLI
Chain SDFYPGAVTVAWKADSSPVKAGVETTTPSK constant
QSNNKYAASSYLSLTPEQWKSHRSYSCQVT region) HEGSTVEKTVAPTECS Nucleic SEQ
ID NO: 119 Acid GGTCAGCCCAAGGCTGCCCCCTCGGTCACT
CTGTTCCCACCCTCGAGTGAGGAGCTTCAA GCCAACAAGGCCACACTGGTGTGTCTCATA
AGTGACTTCTACCCGGGAGCCGTGACAGTG GCCTGGAAGGCAGATAGCAGCCCCGTCAAG
GCGGGAGTGGAGACCACCACACCCTCCAAA CAAAGCAACAACAAGTACGCGGCCAGCAGC
TACCTGAGCCTGACGCCTGAGCAGTGGAAG TCCCACAGAAGCTACAGCTGCCAGGTCACG
CATGAAGGGAGCACCGTGGAGAAGACAGTG GCCCCTACAGAATGTTCA KC Amino SEQ ID
NO: 117 (kappa acid RTVAAPSVFIFPPSDEQLKSGTASVVCLLN Chain
NFYPREAKVQWKVDNALQSGNSQESVTEQD constant
SKDSTYSLSSTLTLSKADYEKHKVYACEVT region) HQGLSSPVTKSFNRGEC Nucleic
SEQ ID NO: 120 Acid CGTACGGTGGCTGCACCATCTGTCTTCATC
TTCCCGCCATCTGATGAGCAGTTGAAATCT GGAACTGCCTCTGTTGTGTGCCTGCTGAAT
AACTTCTACCCCAGAGAAGCCAAAGTGCAG TGGAAGGTGGACAACGCCCTGCAGAGCGGA
AACAGCCAGGAAAGCGTGACAGAGCAGGAT TCCAAGGATTCCACATACAGCCTGAGCAGC
ACACTGACACTGTCCAAGGCCGACTACGAG AAGCACAAGGTGTACGCCTGCGAAGTGACA
CACCAGGGACTGTCCTCCCCTGTGACAAAG AGCTTCAACAGAGGAGAATGC
[0358] In some embodiments, signal peptide may be added when
expressing the antibodies of the present disclosure, these signal
peptides may be partially or full removed by host cells during the
secretion of the antibody. In certain embodiments, for expressing
the 26 exemplary antibodies of the present disclosure, signal
peptide (SEQ ID NO: 130: MGWSCIILFLVATATGVHS) is included when
expressing the heavy chain, signal peptide (SEQ ID NO: 131:
MGWSCIILFLVATATGSWA) is included when expressing the light
chain.
[0359] Table 11 which is appended at the end of the specification
shows sequences and SEQ ID NOs mentioned or used in the present
application.
[0360] Antibody Variants
[0361] In certain embodiments, the antibody or antigen binding
fragments thereof provided herein comprise one or more mutations in
one or more of the CDR sequences provided in Table 1 above, one or
more of the non-CDR sequences of the heavy chain variable region or
light chain variable region provided in Table 2, and/or the
constant region (e.g. Fc region) in Table 4, yet retaining specific
binding affinity to RBD of spike protein of SARS-CoV-2. These are
also referred to as variants of antibodies P2A-1A8, P2A-1A9,
P2B-2G11, P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4, P2C-1A3, P2C-1C8,
P2C-1C10, P2C-1D5, P2C-1F11, P2B-1G5, P2B-1A1, P2C-1D7, P2B-1A10,
P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2,
P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6,
P5A-1B8, P5A-1B9, P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9, P5A-2H3,
P5A- 3A1, P5A-3A6, P5A-3B4, P5A-3C12, P22A-1D1, or the antigen
binding fragments thereof. "Mutations" or "mutated" as used herein
include substitutions, insertions, and/or deletions in an amino
acid sequence or polynucleotide sequence. In certain embodiments,
at least one (or all) of the mutation(s) comprises a conservative
substitution.
[0362] In certain embodiments, the variants comprise 1, 2, or 3 CDR
sequences having at least 80% (e.g. at least 85%, 88%, 90%, 91%,
92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) sequence identity to that
(or those) listed in Table 1 above, and in the meantime retain the
binding specificity to SARS-COV-2, optionally having binding
affinity at a level similar to or even higher than its parent
antibody.
[0363] In certain embodiments, the variants comprise one or more
variable region sequences having at least 80% (e.g. at least 85%,
88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) sequence
identity to that (or those) listed in Table 2 above, and in the
meantime retain the binding specificity to SARS-COV-2, optionally
having binding affinity to SARS-COV-2 at a level similar to or even
higher than its parent antibody. In some embodiments, a total of 1
to 10 amino acids have been mutated in a variable region sequence
listed in Table 2 above. In some embodiments, the mutations occur
in the non-CDR sequences (e.g. in the FRs). In some embodiments,
the mutations are conservative substitutions.
[0364] In certain embodiments, the present disclosure provides a
variant of antibody P2A-1A8, P2A-1A9, P2B-2G11, P2A-1A10, P2A-1B3,
P2B-2F6, P2B-2G4, P2C-1A3, P2C-1C8, P2C-1C10, P2C-1D5, P2C-1F11,
P2B-1G5, P2B-1A1, P2C-1D7, P2B-1A10, P2B-1D9, P2B- 1E4, P2B-1G1,
P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2, P5A-2F11, P5A-2E1, P5A-1C8,
P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6, P5A-1B8, P5A-1B9, P5A-1D1,
P5A-1D10, P5A- 2D11, P5A-2G9, P5A-2H3, P5A-3A1, P5A-3A6, P5A-3B4,
P5A-3C12, or P22A-1D1, wherein the variant comprises:
[0365] a) a heavy chain CDR1 (HCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to a HCDR1 sequence of the parent
antibody listed in Table 1, and/or
[0366] b) a heavy chain CDR2 (HCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to a HCDR2 sequence of the parent
antibody listed in Table 1, and/or
[0367] c) a heavy chain CDR3 (HCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to a HCDR3 sequence of the parent
antibody listed in Table 1, and/or
[0368] d) a light chain CDR1 (LCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to a LCDR1 sequence of the parent
antibody listed in Table 1, and/or
[0369] e) a light chain CDR2 (LCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to a LCDR2 sequence of the parent
antibody listed in Table 1, and/or
[0370] f) a light chain CDR3 (LCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to a LCDR3 sequence of the parent
antibody listed in Table 1, and
in the meantime retain the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than its parent antibody.
[0371] In certain embodiments, the antibody variants provided
herein comprises an HCDR1 having no more than 3, 2, or 1 amino acid
mutations in a HCDR1 sequence of the parent antibody listed in
Table 1, an HCDR2 having no more than 6, 5, 4, 3, 2, or 1 amino
acid mutations in a HCDR2 sequence of the parent antibody listed in
Table 1, HCDR3 having no more than 6, 5, 4, 3, 2, or 1 amino acid
mutations in a HCDR3 sequence of the parent antibody listed in
Table 1, LCDR1 having no more than 2 or 1 amino acid mutations in a
LCDR1 sequence of the parent antibody listed in Table 1, LCDR2
having no more than 3, 2, or 1 amino acid mutations in a LCDR2
sequence of the parent antibody listed in Table 1, and/or LCDR3
having no more than 3, 2, or 1 amino acid mutations in a LCDR3
sequence of the parent antibody listed in Table 1, and in the
meantime retain the binding specificity to SARS-COV-2, optionally
having binding affinity to SARS-COV-2 at a level similar to or even
higher than its parent antibody.
[0372] In certain embodiments, the antibody variants provided
herein comprises:
[0373] a) at least one heavy chain CDR sequence having no more than
3, 2, or 1 amino acid substitutions in a heavy chain CDR sequence
of the parent antibody listed in Table 1, or
[0374] b) at least two heavy chain CDR sequences each having no
more than 3, 2, or 1 amino acid substitutions in a heavy chain CDR
sequence of the parent antibody listed in Table 1, or
[0375] c) three heavy chain CDR sequences each having no more than
3, 2, or 1 amino acid substitutions in a heavy chain CDR sequence
of the parent antibody listed in Table 1, or
[0376] d) at least one light chain sequence having no more than 3,
2, or 1 amino acid substitutions in a heavy chain CDR sequence of
the parent antibody listed in Table 1, or
[0377] e) at least two light chain CDR sequences each having no
more than 3, 2, or 1 amino acid substitutions in a heavy chain CDR
sequence of the parent antibody listed in Table 1, or
[0378] f) three light chain CDR sequences each having no more than
3, 2, or 1 amino acid substitutions in a heavy chain CDR sequence
of the parent antibody listed in Table 1, and
[0379] in the meantime retains the binding specificity to
SARS-COV-2, optionally having binding affinity at a level similar
to or even higher than its parent antibody.
[0380] In certain embodiments, the antibody variants provided
herein retains at least part of (or the entirety of) the paratope
of their parent antibodies. As used herein, the term "paratope"
with respect to an antibody refers to a group of amino acid
residues on the variable regions of the antibody that makes direct
contact with the antigen and form the antigen binding site of the
variable regions. A paratope normally comprises or consists of
amino acid residues in one or more CDR sequences.
[0381] In certain embodiments, the present disclosure provides
variants of antibody P2B-2F6, wherein the variant comprises:
[0382] a) a heavy chain CDR1 (HCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 41, and/or
[0383] b) a heavy chain CDR2 (HCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 42, and/or
[0384] c) a heavy chain CDR3 (HCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 43, and/or
[0385] d) a light chain CDR1 (LCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 44, and/or
[0386] e) a light chain CDR2 (LCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 45, and/or
[0387] f) a light chain CDR3 (LCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 46, and
in the meantime retain the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than antibody P2B-2F6.
[0388] In certain embodiments, the antibody variants of antibody
P2B-2F6 comprises an HCDR1 having no more than 4, 3, 2, or 1 amino
acid mutations in SEQ ID NO: 41, an HCDR2 having no more than 3, 2,
or 1 amino acid mutations in SEQ ID NO: 42, HCDR3 having no more
than 6, 5, 4, 3, 2, or 1 amino acid substitutions in SEQ ID NO: 43,
LCDR1 having no more than 4, 3, 2, or 1 amino acid mutations in SEQ
ID NO: 44, LCDR2 having no more than 3, 2, or 1 amino acid
mutations in SEQ ID NO: 45, and/or LCDR3 having no more than 4, 3,
2, or 1 amino acid mutations in SEQ ID NO: 46, and in the meantime
retain the binding specificity to SARS-COV-2, optionally having
binding affinity to SARS-COV-2 at a level similar to or even higher
than antibody P2B-2F6.
[0389] In certain embodiments, the variants of antibody P2B-2F6
retain the entirety of the paratope of antibody P2B-2F6 while one
or more of the amino acid residues outside the paratope of the
antibody may be mutated. In certain embodiments, the paratope of
antibody P2B-2F6 comprises or consists of: Y27, S28, S30, S31, and
Y33 of HCDR1; H54 of HCDR2; G102, I103, V105, V106 and P107 of
HCDR3; and/or G31, Y32 and N33 of LCDR1; wherein the numbering of
residues in the heavy chain CDRs is according to SEQ ID NO: 47, and
the numbering of residues in the light chain CDR is according to
SEQ ID NO: 48.
[0390] In certain embodiments, the variants of antibody P2B-2F6
retain at least part of the paratope of antibody P2B-2F6. For
example, the variants of antibody P2B-2F6 retain at least 60%, at
least 70%, at least 80%, or at least 90% of the residues of the
paratope of antibody P2B-2F6. In certain embodiments, the variants
of antibody P2B-2F6 comprises one or more mutations (e.g.
conservative substitutions) in the paratope of antibody P2B-2F6. In
certain embodiments, the variants of antibody P2B-2F6 comprises no
more than 5, 4, 3, 2 or 1 mutations (e.g. substitutions) in the
paratope of antibody P2B-2F6. In certain embodiments, the variants
of antibody P2B-2F6 comprises no more than 5, 4, 3, 2 or 1
conservative substitutions in the paratope of antibody P2B-2F6.
[0391] In certain embodiments, the present disclosure provides
variants of antibody P2C-1F11, wherein the variant comprises:
[0392] a) a heavy chain CDR1 (HCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 105, and/or
[0393] b) a heavy chain CDR2 (HCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 106, and/or
[0394] c) a heavy chain CDR3 (HCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 107, and/or
[0395] d) a light chain CDR1 (LCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 108, and/or
[0396] e) a light chain CDR2 (LCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 109, and/or
[0397] f) a light chain CDR3 (LCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 110, and
in the meantime retain the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than antibody P2C-1F11.
[0398] In certain embodiments, the antibody variants of antibody
P2C-1F11 comprises an HCDR1 having no more than 4, 3, 2, or 1 amino
acid mutations in SEQ ID NO: 105, an HCDR2 having no more than 3,
2, or 1 amino acid mutations in SEQ ID NO: 106, HCDR3 having no
more than 6, 5, 4, 3, 2, or 1 amino acid mutations in SEQ ID NO:
107, LCDR1 having no more than 4, 3, 2, or 1 amino acid mutations
in SEQ ID NO: 108, LCDR2 having no more than 3, 2, or 1 amino acid
mutations in SEQ ID NO: 109, and/or LCDR3 having no more than 4, 3,
2, or 1 amino acid mutations in SEQ ID NO: 110, and in the meantime
retain the binding specificity to SARS-COV-2, optionally having
binding affinity to SARS-COV-2 at a level similar to or even higher
than antibody P2C-1F11.
[0399] In certain embodiments, the variants of antibody P2C-1F11
retain the entirety of the paratope of antibody P2C-1F11 while one
or more of the amino acid residues outside the paratope of the
antibody may be mutated. In certain embodiments, the paratope of
antibody P2C-1F11 comprises or consists of: G26, 127, T28, S31, N32
and Y33 of HCDR1; Y52, S53, G54, and S56 of HCDR2; R97, L99, V100,
V101, Y102 and D105 of HCDR3; and/or S28, S30 and Y33 of LCDR1;
wherein the numbering of residues in heavy chain is according to
SEQ ID NO: 111, and the numbering of residues in light chain CDR is
according to SEQ ID NO: 112.
[0400] In certain embodiments, the variants of antibody P2C-1F11
retain at least part of the paratope of antibody P2C-1F11. For
example, the variants of antibody P2C-1F11 retain at least 60%, at
least 70%, at least 80%, or at least 90% of the residues of the
paratope of antibody P2C-1F11. In certain embodiments, the variants
of antibody P2C-1F11 comprises one or more mutations or
substitutions (e.g. conservative substitutions) in the paratope of
antibody P2C-1F11. In certain embodiments, the variants of antibody
P2C-1F11 comprises no more than 6, 5, 4, 3, 2 or 1 mutations (e.g.
substitutions) in the paratope of antibody P2C-1F11. In certain
embodiments, the variants of antibody P2C-1F11 comprises no more
than 6, 5, 4, 3, 2 or 1 conservative substitutions in the paratope
of antibody P2C-1F11.
[0401] In certain embodiments, the present disclosure provides
variants of antibody P22A-1D1, wherein the variant comprises:
[0402] a) a heavy chain CDR1 (HCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 426, and/or
[0403] b) a heavy chain CDR2 (HCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 427, and/or
[0404] c) a heavy chain CDR3 (HCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 428, and/or
[0405] d) a light chain CDR1 (LCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 429, and/or
[0406] e) a light chain CDR2 (LCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 430, and/or
[0407] f) a light chain CDR3 (LCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 431, and
in the meantime retain the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than antibody P22A-1D1.
[0408] In certain embodiments, the antibody variants of antibody
P22A-1D1 comprises an HCDR1 having no more than 6, 5, 4, 3, 2, or 1
amino acid mutations in SEQ ID NO: 426, an HCDR2 having no more
than 5, 4, 3, 2, or 1 amino acid mutations in SEQ ID NO: 427, HCDR3
having no more than 6, 5, 4, 3, 2, or 1 amino acid substitutions in
SEQ ID NO: 428, LCDR1 having no more than 5, 4, 3, 2, or 1 amino
acid mutations in SEQ ID NO: 429, LCDR2 having no more than 3, 2,
or 1 amino acid mutations in SEQ ID NO: 430, and/or LCDR3 having no
more than 4, 3, 2, or 1 amino acid mutations in SEQ ID NO: 431, and
in the meantime retain the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than antibody P22A-1D1.
[0409] In certain embodiments, the variants of antibody P22A-1D1
retain the entirety of the paratope of antibody P22A-1D1 while one
or more of the amino acid residues outside the paratope of the
antibody may be mutated. In certain embodiments, the paratope of
antibody P22A-1D1 comprises or consists of: G26, F27, T28, S31, N32
and Y33 of HCDR1; Y52, S53, G54, and S56 of HCDR2; Y58 of heavy
chain framework region 3, R97, R99, D100, Y101, Y102 and D105 of
HCDR3; Q27, G28, 129, S30 and Y32 of LCDR1; S67 of LCDR2; and/or
H90, L91, N92 and Y94 of LCDR3; wherein the numbering of residues
in the heavy chain CDRs is according to SEQ ID NO: 432, and the
numbering of residues in the light chain CDR is according to SEQ ID
NO: 433.
[0410] In certain embodiments, the variants of antibody P22A-1D1
retain at least part of the paratope of antibody P22A-1D1. For
example, the variants of antibody P22A-1D1 retain at least 60%, at
least 70%, at least 80%, or at least 90% of the residues of the
paratope of antibody P22A-1D1. In certain embodiments, the variants
of antibody P22A-1D1 comprises one or more mutations (e.g.
conservative substitutions) in the paratope of antibody P22A-1D1.
In certain embodiments, the variants of antibody P22A-1D1 comprises
no more than 5, 4, 3, 2 or 1 mutations (e.g. substitutions) in the
paratope of antibody P22A-1D1. In certain embodiments, the variants
of antibody P22A-1D1 comprises no more than 5, 4, 3, 2 or 1
conservative substitutions in the paratope of antibody
P22A-1D1.
[0411] In certain embodiments, the present disclosure provides
variants of antibody P5A-1D2, wherein the variant comprises:
[0412] a) a heavy chain CDR1 (HCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 236, and/or
[0413] b) a heavy chain CDR2 (HCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 237, and/or
[0414] c) a heavy chain CDR3 (HCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 238, and/or
[0415] d) a light chain CDR1 (LCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 239, and/or
[0416] e) a light chain CDR2 (LCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 240, and/or
[0417] f) a light chain CDR3 (LCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 241, and
in the meantime retain the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than antibody P5A-1D2.
[0418] In certain embodiments, the antibody variants of antibody
P5A-1D2 comprises an HCDR1 having no more than 6, 5, 4, 3, 2, or 1
amino acid mutations in SEQ ID NO: 236, an HCDR2 having no more
than 9, 8, 7, 6, 5, 4, 3, 2, or 1 amino acid mutations in SEQ ID
NO: 237, HCDR3 having no more than 6, 5, 4, 3, 2, or 1 amino acid
substitutions in SEQ ID NO: 238, LCDR1 having no more than 4, 3, 2,
or 1 amino acid mutations in SEQ ID NO: 239, LCDR2 having no more
than 4, 3, 2, or 1 amino acid mutations in SEQ ID NO: 240, and/or
LCDR3 having no more than 4, 3, 2, or 1 amino acid mutations in SEQ
ID NO: 241, and in the meantime retain the binding specificity to
SARS-COV-2, optionally having binding affinity to SARS-COV-2 at a
level similar to or even higher than antibody P5A-1D2.
[0419] In certain embodiments, the variants of antibody P5A-1D2
retain the entirety of the paratope of antibody P5A-1D2 while one
or more of the amino acid residues outside the paratope of the
antibody may be mutated. In certain embodiments, the paratope of
antibody P5A-1D2 comprises or consists of: G26, F27, 128, S31, N32
and Y33 of HCDR1; Y52, S53, G54, and S56 of HCDR2; Y58 and R87 of
heavy chain framework region 3, R97, L99, Q100, V101, G102, A103,
T104 and D106 of HCDR3; A31 and Y33 of LCDR1; and/or S95 of LCDR3;
wherein the numbering of residues in the heavy chain CDRs is
according to SEQ ID NO: 242, and the numbering of residues in the
light chain CDR is according to SEQ ID NO: 243.
[0420] In certain embodiments, the variants of antibody P5A-1D2
retain at least part of the paratope of antibody P5A-1D2. For
example, the variants of antibody P5A-1D2 retain at least 60%, at
least 70%, at least 80%, or at least 90% of the residues of the
paratope of antibody P5A-1D2. In certain embodiments, the variants
of antibody P5A-1D2 comprises one or more mutations (e.g.
conservative substitutions) in the paratope of antibody P5A-1D2. In
certain embodiments, the variants of antibody P5A-1D2 comprises no
more than 5, 4, 3, 2 or 1 mutations (e.g. substitutions) in the
paratope of antibody P5A-1D2. In certain embodiments, the variants
of antibody P5A-1D2 comprises no more than 5, 4, 3, 2 or 1
conservative substitutions in the paratope of antibody P5A-1D2.
[0421] In certain embodiments, the present disclosure provides
variants of antibody P5A-3C8, wherein the variant comprises:
[0422] a) a heavy chain CDR1 (HCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 226, and/or
[0423] b) a heavy chain CDR2 (HCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 227, and/or
[0424] c) a heavy chain CDR3 (HCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 228, and/or
[0425] d) a light chain CDR1 (LCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 229, and/or
[0426] e) a light chain CDR2 (LCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 230, and/or
[0427] f) a light chain CDR3 (LCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 231, and
in the meantime retain the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than antibody P5A-3C8.
[0428] In certain embodiments, the antibody variants of antibody
P5A-3C8 comprises an HCDR1 having no more than 6, 5, 4, 3, 2, or 1
amino acid mutations in SEQ ID NO: 226, an HCDR2 having no more
than 5, 4, 3, 2, or 1 amino acid mutations in SEQ ID NO: 227, HCDR3
having no more than 6, 5, 4, 3, 2, or 1 amino acid substitutions in
SEQ ID NO: 228, LCDR1 having no more than 5, 4, 3, 2, or 1 amino
acid mutations in SEQ ID NO: 229, LCDR2 having no more than 3, 2,
or 1 amino acid mutations in SEQ ID NO: 230, and/or LCDR3 having no
more than 5, 4, 3, 2, or 1 amino acid mutations in SEQ ID NO: 231,
and in the meantime retain the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than antibody P5A-3C8.
[0429] In certain embodiments, the variants of antibody P5A-3C8
retain the entirety of the paratope of antibody P5A-3C8 while one
or more of the amino acid residues outside the paratope of the
antibody may be mutated. In certain embodiments, the paratope of
antibody P5A-3C8 comprises or consists of: G26, F27, T28, S31, N32
and Y33 of HCDR1; Y52, S53, G54, and S56 of HCDR2; Y58 of heavy
chain framework region 3, R97, L99, Q100, E101 and H102 of HCDR3;
and G28, 129, S30, S31 and Y32 of LCDR1; S67 of LCDR2; G68 of light
chain framework region 3, H90, L91, N92, S93 and Y94 of LCDR3;
wherein the numbering of residues in the heavy chain CDRs is
according to SEQ ID NO: 232, and the numbering of residues in the
light chain CDR is according to SEQ ID NO: 233.
[0430] In certain embodiments, the variants of antibody P5A-3C8
retain at least part of the paratope of antibody P5A-3C8. For
example, the variants of antibody P5A-3C8 retain at least 60%, at
least 70%, at least 80%, or at least 90% of the residues of the
paratope of antibody P5A-3C8. In certain embodiments, the variants
of antibody P5A-3C8 comprises one or more mutations (e.g.
conservative substitutions) in the paratope of antibody P5A-3C8. In
certain embodiments, the variants of antibody P5A-3C8 comprises no
more than 5, 4, 3, 2 or 1 mutations (e.g. substitutions) in the
paratope of antibody P5A-3C8. In certain embodiments, the variants
of antibody P5A-3C8 comprises no more than 5, 4, 3, 2 or 1
conservative substitutions in the paratope of antibody P5A-3C8.
[0431] The variants of the antibodies or the antigen binding
fragments thereof can retain their parent antibodies' binding
specificity to RBD of the spike protein of SARS-CoV-2, or may
further have one or more desirable properties conferred by the
mutation(s). For example, the variants may have improved
antigen-binding affinity, improved glycosylation pattern, reduced
risk of glycosylation, reduced deamination, reduced or depleted
effector function(s), improved FcRn receptor binding in a pH
dependent manner, increased pharmacokinetic half-life, pH
sensitivity, and/or compatibility to conjugation (e.g. one or more
introduced cysteine residues). Such variants are also known as
affinity variants, glycosylation variants, cysteine variants, Fc
variants, and so on, which are described in more details as
follows.
[0432] a) Affinity Variant
[0433] Affinity variant may contain modifications or substitutions
in one or more CDR sequences as provided in Table 1 above, one or
more framework (FR) sequences provided herein, or the heavy or
light chain variable region sequences provided in Table 2 above. FR
sequences can be readily identified by a skilled person in the art
based on the CDR sequences in Table 1 above and variable region
sequences in Table 2 above, as it is well-known in the art that a
CDR region is flanked by two FR regions in the variable region.
[0434] The affinity variants retain specific binding affinity to
RBD of the spike protein of SARS-COV-2 of the parent antibody, or
even have improved specific binding affinity to the RBD of the
spike protein of SARS-CoV-2 over the parent antibody. Various
methods known in the art can be used to achieve this purpose. For
example, a library of antibody variants (such as Fab or scFv
variants) can be generated and expressed with phage display
technology, and then screened for the binding affinity to the RBD
of the spike protein of SARS-COV-2. For another example, computer
software can be used to virtually simulate the binding of the
antibodies to the RBD of the spike protein of SARS-COV-2, and
identify the amino acid residues on the antibodies which form the
binding interface. Such residues may be either avoided in the
substitution so as to prevent reduction in binding affinity, or
targeted for substitution to provide for a stronger binding.
[0435] In certain embodiments, the affinity variant provided herein
comprises one or more amino acid residue substitutions in one or
more CDR sequences, and/or one or more FR sequences. In certain
embodiments, an affinity variant comprises no more than 20, 15, 10,
9, 8, 7, 6, 5, 4, 3, 2, or 1 substitutions in the CDR sequences
and/or FR sequences in total.
[0436] b) Glycosylation Variant
[0437] The anti-SARS-COV-2 antibodies and antigen-binding fragments
provided herein also encompass a glycosylation variant, which can
be obtained to either increase or decrease the extent of
glycosylation of the antibody or antigen binding fragment
thereof.
[0438] The antibody or antigen binding fragment thereof may
comprise one or more modifications that introduces or removes a
glycosylation site. A glycosylation site is an amino acid residue
with a side chain to which a carbohydrate moiety (e.g. an
oligosaccharide structure) can be attached. Glycosylation of
antibodies is typically either N-linked or O-linked. N-linked
refers to the attachment of the carbohydrate moiety to the side
chain of an asparagine residue, for example, an asparagine residue
in a tripeptide sequence such as asparagine-X-serine and
asparagine-X-threonine, where X is any amino acid except proline.
O-linked glycosylation refers to the attachment of one of the
sugars N-aceylgalactosamine, galactose, or xylose to a hydroxyamino
acid, most commonly to serine or threonine. Removal of a native
glycosylation site can be conveniently accomplished, for example,
by altering the amino acid sequence such that one of the
above-described tripeptide sequences (for N-linked glycosylation
sites) or serine or threonine residues (for O-linked glycosylation
sites) present in the sequence in the is substituted. A new
glycosylation site can be created in a similar way by introducing
such a tripeptide sequence or serine or threonine residue.
[0439] In certain embodiments, the anti-SARS-COV-2 antibodies and
antigen-binding fragments provided herein comprise a mutation at
N297 (e.g. N297A, N297Q, or N297G) to remove the glycosylation
site.
[0440] c) Cysteine-Engineered Variant
[0441] The anti-SARS-COV-2 antibodies and antigen-binding fragments
provided herein also encompass a cysteine-engineered variant, which
comprises one or more introduced free cysteine amino acid
residues.
[0442] A free cysteine residue is one which is not part of a
disulfide bridge. A cysteine-engineered variant is useful for
conjugation with for example, a cytotoxic and/or imaging compound,
a label, or a radioisoptype among others, at the site of the
engineered cysteine, through for example a maleimide or haloacetyl.
Methods for engineering antibodies or antigen-binding fragments
thereof to introduce free cysteine residues are known in the art,
see, for example, WO2006/034488.
[0443] d) Fc Variant
[0444] The anti-SARS-COV-2 antibodies and antigen-binding fragments
provided herein also encompass an Fc variant, which comprises one
or more amino acid residue modifications or substitutions at its Fc
region and/or hinge region, for example, to provide for altered
effector functions such as ADCC, ADCP and CDC. Methods of altering
ADCC activity by antibody engineering have been described in the
art, see for example, Shields R L. et al., J Biol Chem. 2001.
276(9): 6591-604; Idusogie E E. et al., J Immunol.
2000.164(8):4178-84; Steurer W. et al., J Immunol. 1995, 155(3):
1165-74; Idusogie E E. et al., J Immunol. 2001, 166(4): 2571-5;
Lazar G A. et al., PNAS, 2006, 103(11): 4005-4010; Ryan M C. et
al., Mol. Cancer Ther., 2007, 6: 3009-3018; Richards J O., et al.,
Mol Cancer Ther. 2008, 7(8): 2517-27; Shields R. L. et al, J. Biol.
Chem, 2002, 277: 26733-26740; Shinkawa T. et al, J. Biol. Chem,
2003, 278: 3466-3473.
[0445] CDC activity of the antibodies provided herein can also be
altered, for example, by improving or diminishing C1q binding
and/or CDC (see, for example, WO99/51642; Duncan & Winter
Nature 322:738-40 (1988); U.S. Pat. Nos. 5,648,260; 5,624,821); and
WO94/29351 concerning other examples of Fc region variants. One or
more amino acids selected from amino acid residues 329, 331 and 322
of the Fc region can be replaced with a different amino acid
residue to alter C1q binding and/or reduced or abolished complement
dependent cytotoxicity (CDC) (see, U.S. Pat. No. 6,194,551 by
Idusogie et al). One or more amino acid substitution(s) can also be
introduced to alter the ability of the antibody to fix complement
(see PCT Publication WO 94/29351 by Bodmer et al.).
[0446] The term "Antibody-dependent cellular phagocytosis" and
"ADCP" refer to a process by which antibody-coated cells or
particles are internalized, either in whole or in part, by
phagocytic immune cells (e.g., macrophages, neutrophils and
dendritic cells) that bind to an immunoglobulin Fc region. Methods
for altering the ADCP activity of antibodies by antibody
engineering are known in the art, see for example, Kellner C et
al., Transfus Med Hemother, (2017)44:327-336 and Chung A W et al.,
AIDS, (2014) 28:2523-2530.
[0447] Examples of Fc variants are known in the art, see, for
example, Wang et al., Protein Cell 2018, 9(1): 63-73 and Kang et
al., Exp & Mol., Med. (2019) 51:138, which are incorporated
herein to their entirety.
[0448] i) Fc Variant with Enhanced Effector Functions
[0449] In certain embodiments, the Fc variants provided herein has
increased ADCC and/or increased affinity to an Fc.gamma. receptor
(e.g. Fc.gamma.RI (CD64), Fc.gamma.RII (CD32) and/or Fc.gamma.RIII
(CD16)) relative to a wildtype Fc (e.g. Fc of IgG1). In certain
embodiments, an Fc variant comprises one or more amino acid
substitution(s) at one or more of the following positions: 234,
235, 236, 238, 239, 240, 241, 243, 244, 245, 246, 247, 248, 249,
252, 254, 255, 256, 258, 260, 262, 263, 264, 265, 267, 268, 269,
270, 272, 274, 276, 278, 280, 283, 285, 286, 289, 290, 292, 293,
294, 295, 296, 298, 299, 300, 301, 303, 304, 305, 307, 309, 312,
313, 315, 320, 322, 324, 325, 326, 327, 329, 330, 331, 332, 333,
334, 335, 337, 338, 339, 340, 345, 360, 373, 376, 378, 382, 388,
389, 396, 398, 414, 416, 419, 430, 433, 434, 435, 436, 437, 438,
439 and 440 of the Fc region (see WO 00/42072 by Presta,
WO2006/019447 by Lazar, and WO2016/196228, incorporated herein to
its entirety), wherein the numbering of the residues in the Fc
region is that of the EU index as in Kabat (see, Kabat E. A. et
al., Sequences of Proteins of immunological Interest, 5th Ed.
Public Health Service, National Institutes of Health, Bethesda, Md.
(1991)). Exemplary substitutions for increased effector functions
include, without limitation, 234Y, 235Q, 236A, 236W, 239D, 239E,
239M, 243L, 247I, 268D, 267E, 268D, 268E, 268F, 270E, 280H, 290S,
292P, 298A, 298D, 298V, 300L, 305I, 324T, 326A, 326D, 326W, 330L,
330M, 333S, 332D, 332E, 298A, 333A, 334A, 334E, 326A, 247I, 339D,
339Q, 345R, 280H, 290S, 298D, 298V, 243L, 292P, 300L, 396L, 305I,
396L, 430G, 440Y, or any combination thereof (such as 239D/332E,
239D/332E/330L, 236A/332E, 236A/239D/332E, 268F/324T, 267E/268F,
267E/324T, and 267E/268F/324T) (see, WO2016/196228; Richards et al.
(2008) Mol. Cancer Therap. 7:2517; Moore et al. (2010) mAbs 2:181;
and Strohl (2009) Current Opinion in Biotechnology 20:685-691).
[0450] Specific mutations at positions 256, 290, 298, 333, 334 and
339 were shown to improve binding to Fc.gamma.RIII. Additionally,
the following combination mutants were shown to improve
Fc.gamma.RIII binding: T256A/S298A, S298A/E333A, S298A/K224A,
F243L/R292P/Y300L/V305I/P396L, S298A/E333A/K334A and
L234Y/L235Q/G236W/S239M/H268D/D270E/S298A in one heavy chain and
D270E/K326D/A330M/K334E in the opposing heavy chain (having
enhanced Fc.gamma.RIII binding and ADCC activity). Other Fc
variants with strongly enhanced binding to Fc.gamma.RIIIa include
variant with S239D/I332E and S239D/I332E/A330L mutations, which
showed the greatest increase in affinity for Fc.gamma.RIIIa, a
decrease in Fc.gamma.RIIb binding, and strong cytotoxic activity,
and variants with L235V, F243L, R292P, Y300L, V305I and P396L
mutations, which exhibited enhancing Fc.gamma.RIIIa and
concomitantly enhanced ADCC activity. (see Lazar et a. (2006) Proc.
Nat'l Acad Sci. (USA) 103:4005; Awan et al. (2010) Blood 115: 1204;
Desjarlais & Lazar (2011) Exp. Cell Res, Stavenhagen et al.
(2007) Cancer Res 67:8882). Modifications that increase binding to
Clq can be introduced in order to enhance CDC activity. Exemplary
modifications include, a K326 (e.g., K326W) and/or E333
modification in an IgG2, or a S267E/H268F/S324T modification, alone
or in any combination, in an IgGl (see Idusogie et al. (2001) J.
Immunol. 166:2571, Moore et al. (2010) mAbs 2: 181). Other
exemplary modifications include, K326W/E333S, S267E/H268F/S324T,
and E345R/E430G/S440Y.
[0451] ii) Fc with Reduced Effector Functions
[0452] In certain embodiments, the Fc variants provided herein has
reduced effector functions relative to a wildtype Fc (e.g. Fc of
IgG1), and comprise one or more amino acid substitution(s) at a
position selected from the group consisting of: 220, 226, 229, 233,
234, 235, 236, 237, 238, 267, 268, 269, 270, 297, 309, 318, 320,
322, 325, 328, 329, 330, and 331 of the Fc region (see,
WO2016/196228; Richards et al. (2008) Mol. Cancer Therap. 7:2517;
Moore et al. (2010) mAbs 2:181; and Strohl (2009) Current Opinion
in Biotechnology 20:685-691), wherein the numbering of the residues
in the Fc region is that of the EU index as in Kabat. Exemplary
substitutions for reduced effector functions include, without
limitation, 220S, 226S, 228P, 229S, 233P, 234V, 234G, 234A, 234F,
234A, 235A, 235G, 235E, 236E, 236R, 237A, 237K, 238S, 267R, 268A,
268Q, 269R, 297A, 297Q, 297G, 309L, 318A, 322A, 325L, 328R, 330S,
331S, or any combination thereof (see, WO2016/196228; and Strohl
(2009) Current Opinion in Biotechnology 20:685-691).
[0453] In certain embodiments, the Fc variant provided herein is of
IgG1 isotype and comprises one or more amino acid substitution(s)
selected from the group consisting of: L234A, L234F, L234V, F234A,
V234A, L235A, L235E, G237A, P238S, H268Q, H268A, N297A, N297Q,
N297G, V309L, A330S, and P331S, or any combination thereof (such as
L234A/L235A). In certain embodiments, the Fc variant provided
herein is of IgG2 isotype, and comprises one or more amino acid
substitution(s) selected from the group consisting of: H268Q,
V309L, A330S, P331S, V234A, G237A, P238S, H268A, and any
combination thereof. In certain embodiments, the Fc variant
provided herein is of IgG4 isotype, and comprises one or more amino
acid substitution(s) selected from the group consisting of: S228P,
F234A, L235E, L235A, G237A, E318A, N297A, N297Q, N297G, and any
combination thereof. In certain embodiments, the anti-SARS-COV-2
antibodies and antigen-binding fragments provided herein is of
IgG2/IgG4 cross isotype. Examples of IgG2/IgG4 cross isotype is
described in Rother R P et al, Nat Biotechnol 25:1256-1264
(2007).
[0454] iii) Fc with Altered Binding to FcRn
[0455] In certain embodiments, the Fc variant comprises one or more
amino acid substitution(s) that improves binding affinity to
neonatal Fc receptor (FcRn) at pH 6.0 while retaining minimal
binding at pH 7.4. Such a variant can have an extended
pharmacokinetic half-life, as it binds to FcRn at acidic pH which
allows it to escape from degradation in the lysosome and then be
translocated and released out of the cell. Methods of engineering
an antibody and antigen-binding fragment thereof to improve binding
affinity with FcRn are well-known in the art, see, for example,
Vaughn, D. et al, Structure, 6(1): 63-73, 1998; Kontermann, R. et
al, Antibody Engineering, Volume 1, Chapter 27: Engineering of the
Fc region for improved PK, published by Springer, 2010; Yeung, Y.
et al, Cancer Research, 70: 3269-3277 (2010); Hinton, P. et al, J.
Immunology, 176:346-356 (2006); Petkova et al. (2006) Int. Immunol.
18:1759, Ball Acqua et al. Journal of Immunology 2002,
169:5171-5180, Dall'Acqua W F. et al., J Biol Chem. 281:23514-23524
(2006); Zalevsky J, et al, Nat Biotechnol.; 28:157-159 (2010); WO
2009/086320; U.S. Pat. Nos. 6,277,375; 6,821,505; WO 97/34631; and
WO 2002/060919.
[0456] Non-limiting examples of Fc modifications that may result in
an increase in serum half-life of the antibody when administered
include, e.g., substitution(s) at one or more positions selected
from: 234 (e.g., with F), 235 (e.g., with Q), 238 (e.g., with D),
250 (e.g., with E or Q), 252 (e.g., with L/Y/F/W or T), 254 (e.g.,
with S or T), 256 (e.g., with S/R/Q/E/D or T); 259 (e.g., with I);
272 (e.g., with A), 305(e.g., with A), 307(e.g., with A or P), 308
(e.g., with F, C or P), 311 (e.g., with A or R), 312 (e.g., with
A), 322 (e.g., Q), 328 (e.g. E), 331 (e.g., with A), 378 (e.g.,
with A), 380 (e.g., with A), 382 (e.g., with A), 428 (e.g., with L
or F), 432 (e.g., with C), 433 (e.g., with H/L/R/S/P/Q or K), 434
(e.g., with H/F or Y or S or A or W), 435 (e.g. with H), 436 (e.g.,
with L) and 437 (e.g., with C) (all positions by EU numbering)
(see, WO2016049000A2; WO2020052692; WO2016196228). In some
embodiments, the Fc variant comprises one or more amino acid
substitution(s) selected from the group consisting of 234F, 235Q,
238D, 250Q, 252T, 252Y, 254T, 256E, 259I, 272A, 305A, 307A, 308F,
311A, 322Q, 328E, 331S, 380A, 428L, 432C, 433K, 433S, 434S, 434Y,
434F, 434W, 434A, 435H, 436L, 437C and any combination thereof. In
some embodiments, the Fc modifications comprises one or pairs or
groups of modifications selected from: a) a 428L (e.g., M428L) and
434S (e.g., N434S) substitution; a 428L, 259I (e.g., V259I), and
308F (e.g., V308F) substitution; b) a 433K (e.g., H433K) and 434
(e.g., N434Y or N434F) substitution; c) a 252Y, 254T, and 256E
(e.g., M252Y, S254T, and T256E) substitution; d) a 250Q and 428L
substitution (e.g., T250Q and M428L); e) a 307A, 380A and 434A
substitution (e.g., T307A, E380A and N434A); f) a P238D and L328E
substitution; g) a L234F, L235Q, K322Q, M252T, S254T and T256E
substitution; and h) and a L432C, H433S, N434W, Y436L and T437C
substitution.
[0457] In some embodiments, hybrid IgG isotypes may be used to
increase FcRn binding and half-life of antibodies. A hybrid Ig can
be generated from two or more isotypes. For example, an IgGl/IgG3
hybrid variant may be constructed by substituting IgGl positions in
the CH2 and/or CH3 region with the amino acids from IgG3 at
positions where the two isotypes differ. In some embodiments, a
hybrid Ig can comprises one or more modifications (e.g.
substitutions) disclosed here.
[0458] Antigen-Binding Fragments
[0459] Provided herein are also anti-SARS-CoV-2 antigen-binding
fragments. In some embodiments, the antibodies and antigen-binding
fragments provided herein comprise all or a portion of the heavy
chain variable domain and/or all or a portion of the light chain
variable domain.
[0460] Various types of antigen-binding fragments are known in the
art and can be developed based on the anti-SARS-CoV-2 antibodies
provided herein, including for example, the exemplary antibodies
whose CDR are shown in Tables 1 above, and variable sequences are
shown in Tables 2 and 3, and their different variants (such as
affinity variants, glycosylation variants, Fc variants,
cysteine-engineered variants and so on).
[0461] In certain embodiments, an anti-SARS-CoV-2 antigen-binding
fragment provided herein is a diabody, a Fab, a Fab', a
F(ab').sub.2, a Fd, an Fv fragment, a disulfide stabilized Fv
fragment (dsFv), a (dsFv).sub.2, a bispecific dsFv (dsFv-dsFv'), a
disulfide stabilized diabody (ds diabody), a single-chain antibody
molecule (scFv), an scFv dimer (bivalent diabody), a bispecific
scFv dimer, a multispecific antibody, a heavy chain antibody, a
camelized single domain antibody, a nanobody, a domain antibody,
and a bivalent domain antibody.
[0462] Various techniques can be used for the production of such
antigen-binding fragments. Illustrative methods include, enzymatic
digestion of intact antibodies (see, e.g. Morimoto et al., Journal
of Biochemical and Biophysical Methods 24:107-117 (1992); and
Brennan et al., Science, 229:81 (1985)), recombinant expression by
host cells such as E. coli (e.g. for Fab, Fv and ScFv antibody
fragments), screening from a phage display library as discussed
above (e.g. for ScFv), and chemical coupling of two Fab'-SH
fragments to form F(ab').sub.2 fragments (Carter et al.,
Bio/Technology 10:163-167 (1992)). Other techniques for the
production of antibody fragments will be apparent to a person
skilled in the art.
[0463] In certain embodiments, the antigen-binding fragment is a
scFv. Generation of scFv is described in, for example, WO 93/16185;
U.S. Pat. Nos. 5,571,894; and 5,587,458. ScFv may be fused to an
effector protein at either the amino or the carboxyl terminus to
provide for a fusion protein (see, for example, Antibody
Engineering, ed. Borrebaeck).
[0464] In certain embodiments, the anti-SARS-CoV-2 antibodies or
antigen-binding fragments thereof provided herein are bivalent,
tetravalent, hexavalent, or multivalent. Any molecule being more
than bivalent is considered multivalent, encompassing for example,
trivalent, tetravalent, hexavalent, and so on.
[0465] A bivalent molecule can be monospecific if the two binding
sites are both specific for binding to the same antigen or the same
epitope. This, in certain embodiments, provides for stronger
binding to the antigen or the epitope than a monovalent
counterpart. Similar, a multivalent molecule may also be
monospecific. In certain embodiments, in a bivalent or multivalent
antigen-binding moiety, the first valent of binding site and the
second valent of binding site are structurally identical (i.e.
having the same sequences), or structurally different (i.e. having
different sequences albeit with the same specificity).
[0466] A bivalent can also be bispecific, if the two binding sites
are specific for different or overlapping antigens or epitopes.
This also applies to a multivalent molecule. For example, a
trivalent molecule can be bispecific when two binding sites are
monospecific for a first antigen (or epitope) and the third binding
site is specific for a second antigen (or epitope).
[0467] Bispecific (or Bivalent) Antibody or Antigen-Binding
Fragments
[0468] In another aspect, the present disclosure provides
bispecific (or bivalent) antibody molecules comprising an
anti-SARS-CoV-2 antibody or antigen-binding fragment thereof as
disclosed herein. In certain embodiments, the bispecific (or
bivalent) antibodies provided herein comprises a first
antigen-binding domain and a second antigen-binding domain, wherein
the first antigen-binding domains is derived from a monoclonal
antibody selected from the group consisting of P2A-1A8, P2A-1A9,
P2B-2G11, P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4, P2C-1A3, P2C-1C8,
P2C-1C10, P2C-1D5, P2C-1F11, P2B-1G5, P2B-1A1, P2C-1D7, P2B-1A10,
P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2,
P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6,
P5A-1B8, P5A-1B9, P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9, P5A-2H3,
P5A-3A1, P5A-3A6, P5A-3B4, P5A- 3C12, and P22A-1D1. The second
antigen-binding domain can be derived from any suitable
antibody.
[0469] In certain embodiments, the bispecific (or bivalent)
antibodies provided herein comprises a first antigen-binding domain
and a second antigen-binding domain, wherein the first and the
second antigen-binding domains are derived from any two monoclonal
antibodies selected from the group consisting of P2A-1A8, P2A-1A9,
P2B-2G11, P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4, P2C-1A3, P2C-1C8,
P2C-1C10, P2C-1D5, P2C-1F11, P2B-1G5, P2B- 1A1, P2C-1D7, P2B-1A10,
P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2,
P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6,
P5A- 1B8, P5A-1B9, P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9, P5A-2H3,
P5A-3A1, P5A-3A6, P5A-3B4, P5A-3C12, and P22A-1D1. Any two
monoclonal antibodies from the above 42 antibodies can be combined,
as if each and every possible combination of two antibodies have
been set forth herein individually. In certain embodiments, the
bispecific (or bivalent) antibodies provided herein comprises a
first antigen-binding domain and a second antigen-binding domain,
wherein the first and the second antigen-binding domains are
derived from any two monoclonal antibodies selected from the group
consisting of P2B-2F6, P2C-1F11, P2B-1G5, P2B-1A1, P2C-1D7,
P2B-1A10, P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7, P5A-3C8,
P5A-1D2, P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6, P4B-1F4,
P5A-1B6, P5A-1B8, P5A-1B9, P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9,
P5A-2H3, P5A- 3A1, P5A-3A6, P5A-3B4, P5A-3C12, and P22A-1D1. Any
two monoclonal antibodies from the above 32 antibodies can be
combined, as if each and every possible combination of two
antibodies have been set forth herein individually.
[0470] In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2B-2F6,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2A-1A8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2A-1A9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2B-2G11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2A-1A10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2A-1B3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2B-2G4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2C-1A3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2C-1C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2C-1C10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2C-1D5,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1F11 and P2C-1F11,
respectively.
[0471] In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A8 and P2A-1A9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A8 and P2B-2G11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A8 and P2A-1A10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A8 and P2A-1B3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A8 and P2B-2F6,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A8 and P2B-2G4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A8 and P2C-1A3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A8 and P2C-1C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A8 and P2C-1C10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A8 and P2C-1D5,
respectively.
[0472] In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A9 and 2B-2G11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A9 and P2A-1A10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A9 and P2A-1B3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A9 and P2B-2F6,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A9 and P2B-2G4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A9 and P2C-1A3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A9 and P2C-1C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A9 and P2C-1C10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A9 and P2C-1D5,
respectively.
[0473] In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G11 and P2A-1A10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G11 and P2A-1B3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G11 and P2B-2F6,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G11 and P2B-2G4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G11 and P2C-1A3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G11 and P2C-1C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G11 and P2C-1C10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G11 and P2C-1D5,
respectively.
[0474] In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A10 and P2A-1B3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A10 and P2B-2F6,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A10 and P2B-2G4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A10 and P2C-1A3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A10 and P2C-1C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A10 and P2C-1C10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1A10 and P2C-1D5,
respectively.
[0475] In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1B3 and P2B-2F6,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1B3 and P2B-2G4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1B3 and P2C-1A3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1B3 and P2C-1C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1B3 and P2C-1C10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2A-1B3 and P2C-1D5,
respectively.
[0476] In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2F6 and P2B-2G4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2F6 and P2C-1A3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2F6 and P2C-1C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2F6 and P2C-1C10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2F6 and P2C-1D5,
respectively.
[0477] In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G4 and P2C-1A3,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G4 and P2C-1C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G4 and P2C-1C10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2B-2G4 and P2C-1D5,
respectively.
[0478] In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1A3 and P2C-1C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1A3 and P2C-1C10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1A3 and P2C-1D5,
respectively.
[0479] In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1C8 and P2C-1C10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived from P2C-1C8 and P2C-1D5,
respectively.
[0480] In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1C10 and P2C-1D5,
respectively.
[0481] In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P2B-1G5,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P2B-1A1
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P2C-1D7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P2B-1A10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P2B-1D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P2B-1E4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P2B-1G1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P4A-2D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P5A-2G7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P5A-3C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P5A-1D2,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1F11 and P5A-1C8,
respectively.
[0482] In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P2B-1G5,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P2B-1A1
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P2C-1D7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P2B-1A10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P2B-1D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P2B-1E4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P2B-1G1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P4A-2D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P5A-2G7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P5A-3C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P5A-1D2,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-2F6 and P5A-1C8,
respectively.
[0483] In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P2B-1A1
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P2C-1D7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P2B-1A10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P2B-1D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P2B-1E4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P2B-1G1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P4A-2D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P5A-2G7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P5A-3C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P5A-1D2,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G5 and P5A-1C8,
respectively.
[0484] In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P2C-1D7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P2B-1A10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P2B-1D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P2B-1E4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P2B-1G1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P4A-2D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P5A-2G7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P5A-3C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P5A-1D2,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A1 and P5A-1C8,
respectively.
[0485] In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1D7 and P2B-1A10,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1D7 and P2B-1D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1D7 and P2B-1E4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1D7 and P2B-1G1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1D7 and P4A-2D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1D7 and P5A-2G7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1D7 and P5A-3C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1D7 and P5A-1D2,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1D7 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1D7 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2C-1D7 and P5A-1C8,
respectively.
[0486] In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A10 and P2B-1D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A10 and P2B-1E4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A10 and P2B-1G1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A10 and P4A-2D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A10 and P5A-2G7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A10 and P5A-3C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A10 and P5A-1D2,
respectively.
[0487] In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A10 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A10 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1A10 and P5A-1C8,
respectively.
[0488] In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1D9 and P2B-1E4,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1D9 and P2B-1G1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1D9 and P4A-2D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1D9 and P5A-2G7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1D9 and P5A-3C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1D9 and P5A-1D2,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1D9 and P5A-2F 11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1D9 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1D9 and P5A-1C8,
respectively.
[0489] In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1E4 and P2B-1G1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1E4 and P4A-2D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1E4 and P5A-2G7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1E4 and P5A-3C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1E4 and P5A-1D2,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1E4 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1E4 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1E4 and P5A-1C8,
respectively.
[0490] In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G1 and P4A-2D9,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G1 and P5A-2G7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G1 and P5A-3C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G1 and P5A-1D2,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G1 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G1 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P2B-1G1 and P5A-1C8,
respectively.
[0491] In certain embodiments, the first and the second
antigen-binding domains are derived or from P4A-2D9 and P5A-2G7,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P4A-2D9 and P5A-3C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P4A-2D9 and P5A-1D2,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P4A-2D9 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P4A-2D9 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P4A-2D9 and P5A-1C8,
respectively.
[0492] In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-2G7 and P5A-3C8,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-2G7 and P5A-1D2,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-2G7 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-2G7 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-2G7 and P5A-1C8,
respectively.
[0493] In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-3C8 and P5A-1D2,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-3C8 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-3C8 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-3C8 and P5A-1C8,
respectively.
[0494] In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-1D2 and P5A-2F11,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-1D2 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-1D2 and P5A-1C8,
respectively.
[0495] In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-2F11 and P5A-2E1,
respectively. In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-2F11 and P5A-1C8,
respectively.
[0496] In certain embodiments, the first and the second
antigen-binding domains are derived or from P5A-2E1 and P5A-1C8,
respectively.
[0497] In certain embodiments, the bispecific antibody molecules
can have at least two distinct antigen-binding sites with different
specificities. In certain embodiments, the bispecific antibody
molecules provided herein are capable of binding to different
epitopes on the spike protein of SARS-CoV-2 virus. In some
embodiments, the bispecific antibody molecules provided herein
comprises antigen-binding fragments derived from two or more
antibodies provided herein. In some embodiments, the two or more
antibodies bind to different epitopes in RBD of spike protein of
SARS-CoV-2. In some embodiments, the two or more antibodies are no
more than 70% (or no more than 60%, or no more than 50%)
competitive against each other in binding to RBD of spike protein
of SARS-CoV-2 virus. In certain embodiments, the bispecific
antibody comprises a first antigen-binding domain derived from
P2C-1F11 and a second antigen-binding domain derived from an
antibody selected from the group consisting of P2C-1A3, P2C-1C10,
P2B-2F6, P2B-1G5, and P2A-1B3. In certain embodiments, the
bispecific antibody comprises a first antigen-binding domain
derived from P2C-1A3 and a second antigen-binding domain derived
from an antibody selected from the group consisting of P2C-1F11,
and P2A-1B3. In certain embodiments, the bispecific antibody
comprises a first antigen-binding domain derived from P2B-2F6 and a
second antigen-binding domain derived from an antibody selected
from the group consisting of P2C-1C10, P2C-1F11, P2B-1G5, and
P2A-1B3. In certain embodiments, the bispecific antibody comprises
a first antigen-binding domain derived from P2A-1B3 and a second
antigen-binding domain derived from an antibody selected from the
group consisting of P2C-1A3, P2C-1C10, P2C-1F11, P2B-2F6, and
P2A-1A10. In some embodiments, the two or more antibodies comprise
a first antibody which comprises P2C-1C10 or an antigen binding
fragment thereof, and a second antibody selected from the group
consisting of P2C-1A3, P2C-1F11, and P2A-1B3, or an antigen binding
fragment thereof.
[0498] The term "derived from" as used herein with respect to
antigen-binding domain, means that the antigen-binding domain
comprise at least one heavy chain CDR sequence (e.g.
[0499] comprising heavy chain CDR3, or three heavy chain CDRs) or
at least one light chain CDR sequence (e.g. comprising light chain
CDR3, or three heavy chain CDRs) of the specified monoclonal
antibody. In certain embodiments, the first and the second
antigen-binding domains comprises the heavy chain CDR sequences of
the specified monoclonal antibodies, and/or the light chain CDR
sequences of the specified monoclonal antibodies. In certain
embodiments, the first and the second antigen-binding domains
comprises the heavy chain variable region sequences of the
specified monoclonal antibodies, and/or the light chain variable
region sequences of the specified monoclonal antibodies. All the
CDR sequences and variable region sequences of the specific
monoclonal antibodies are provided in Tables 1 and 2 of the present
disclosure.
[0500] In certain embodiments, the bispecific antibody molecules
provided herein has a first antigen-binding domains specificity
directed to the RBD of the spike protein of SARS-CoV-2 virus and a
second antigen-binding domains specificity directed to a second
antigen. In certain embodiments, the second antigen can be for
example, an epitope outside of RBD on the spike protein of
SARS-CoV-2, S2 protein (i.e. which is cleaved from the spike
protein), nucleocapsid protein of SARS-CoV-2, or alternatively the
second antigen can be an antigen on human immune cells such as T
cell, macrophage cell, natural killer cells, or antigen-presenting
cells.
[0501] In certain embodiments, the bispecific antibody molecules as
provided herein are based on the format of a "whole" antibody, such
as whole IgG or IgG-like molecules. Examples of such bispecific
antibody include but are not limited to, those produced by a
quadroma cell line. In another embodiment, a bispecific IgG-like
molecule can be an appended IgG, which is engineered by appending
either the amino or carboxyl termini of either light or heavy
chains of an IgG of a first specificity with additional
antigen-binding units of a second specificity. The appended
antigen-binding units can be, for example, single domain antibodies
(e.g. unpaired VL or VH, or VHH (i.e. heavy chain variable domain
of a heavy chain antibody)), paired antibody variable domains (e.g.
Fv or scFv) or engineered protein scaffolds. Examples of appended
IgG include, without limitation, Double-variable domain (DVD)-Ig,
which has a second heavy chain variable domain (VH) fused to the VH
of a first heavy chain and a second variable light chain domain
(VL) fused to a first light chain of the IgG. A DVD-Ig can be
bispecific when the first VH/VL and the second VH/VL are selected
to bind to two different antigens. In certain embodiments, a
bispecific IgG or IgG-like molecules can be monovalent for each
antigen and can be produced by co-expression of the two light and
two heavy chains in a single host cell.
[0502] In certain embodiments, the bispecific antibody molecules as
provided herein can be small recombinant bispecific formats based
on variable domains, such as single domain antibody, Fv, and Fab,
which may lack some or all of the antibody constant domains.
Examples of small recombinant bispecific formats include, without
limitation, tandem single chain variable fragment molecules
(taFvs), diabodies (Dbs), single chain diabodies (scDbs) and
various other derivatives of these (see, bispecific antibody
formats as described by Byrne H. et al. (2013) Trends Biotech, 31
(11): 621-632, BiTE (bispecific T cell engager), DARTs, and TandAb.
In certain embodiments, the two antigen-binding moieties can be
linked by a peptide linker.
[0503] In certain embodiments, the bispecific antibody molecules as
provided herein are in a bispecific format selected from bispecific
IgG-like antibodies (BsIgG) comprising CrossMab; DAF (two-in-one);
DAF (four-in-one); DutaMab; DT-IgG; Knobs-in-holes common LC;
Knobs-in-holes assembly; Charge pair; Fab-arm exchange; SEEDbody;
Triomab; LUZ-Y; Fcab; kappa-lamda-body; and Orthogonal Fab. For
detailed description of the bispecific antibody formats please see
Spiess C., Zhai Q. and Carter P. J. (2015) Molecular Immunology 67:
95-106, which is incorporated herein by reference to its
entirety.
[0504] In certain embodiments, the bispecific antibody molecules as
provided herein are in a bispecific format selected from
IgG-appended antibodies with an additional antigen-binding moiety
consisting of DVD-IgG; IgG(H)-scFv; scFv-(H)IgG; IgG(L)-scFv;
scFV-(L)IgG; IgG(L,H)-Fv; IgG(H)-V; V(H)-IgG; IgG(L)-V; V(L)-IgG;
IgG-scFab; 2scFv-IgG; IgG-2scFv; scFv4-Ig; scFv4-Ig; and Zybody
(see Id.).
[0505] In certain embodiments, the bispecific antibody molecules as
provided herein are in a bispecific format selected from WuxiBody
(WuXi Biologics, see, WO2019057122A1, incorporated herein to its
entirety); Triomabs; hybrid hybridoma (quadroma); Multispecific
anticalin platform (Pieris); Diabodies; Single chain diabodies;
Tandem single chain Fv fragments; TandAbs, Trispecific Abs
(Affimed); Darts (dual affinity retargeting; Macrogenics);
Bispecific Xmabs (Xencor); Bispecific T cell engagers (Bites;
Amgen; 55 kDa); Triplebodies; Tribody (Fab-scFv); Fusion Protein
(CreativeBiolabs); multifunctional recombinant antibody derivates;
Duobody platform (Genmab); Dock and lock platform; Knob into hole
(KIH) platform; Humanized bispecific IgG antibody (REGN1979)
(Regeneron); Mabe bispecific antibodies (F-Star); DVD-Ig (dual
variable domain immunoglobulin) (Abbvie); kappa-lambda bodies; TBTI
(tetravalent bispecific tandem Ig); and CrossMab.
[0506] In certain embodiments, the bispecific antibody molecules as
provided herein are in a format selected from bispecific antibody
fragments comprising Nanobody; Nanobody-HAS; BiTE; Diabody; DART;
TandAb; scDiabody; sc-Diabody-CH3; Diabody-CH3; Triple Body;
Miniantibody; Minibody; TriBi minibody; scFv-CH3 KIH; Fab-scFv;
scFv-CH-CL-scFv; F(ab')2; F(ab')2-scFv2; scFv-KIH; Fab-scFv-Fc;
Tetravalent HCAb; scDiabody-Fc; Diabody-Fc; Tandem scFv-Fc; and
Intrabody (see Id.).
[0507] In certain embodiments, the bispecific antibody molecules as
provided herein are in a bispecific format such as Dock and Lock;
ImmTAC; HSAbody; scDiabody-HAS; and Tandem scFv-Toxin (see
Id.).
[0508] In certain embodiments, the bispecific antibody molecules as
provided herein are based on a format selected from bispecific
antibody conjugates comprising IgG-IgG; Cov-X-Body; and
scFv1-PEG-scFv2 (see Id.).
[0509] The bispecific antibody molecules provided herein can be
made with any suitable methods known in the art. In a conventional
approach, two immunoglobulin heavy chain-light chain pairs having
different antigen-binding specificities can be co-expressed in a
host cell to produce bispecific antibodies in a recombinant way
(see, for example, Milstein and Cuello, Nature, 305: 537 (1983)),
followed by purification by affinity chromatography.
[0510] Recombinant approach may also be used, where sequences
encoding the antibody heavy chain variable domains for the two
specificities are respectively fused to immunoglobulin constant
domain sequences, followed by insertion to an expression vector
which is co-transfected with an expression vector for the light
chain sequences to a suitable host cell for recombinant expression
of the bispecific antibody (see, for example, WO 94/04690; Suresh
et al., Methods in Enzymology, 121:210 (1986)). Similarly, scFv
dimers can also be recombinantly constructed and expressed from a
host cell (see, e.g. Gruber et al., J. Immunol., 152:5368
(1994).)
[0511] Bispecific antibody molecule may be generated from a
bispecific antibody, for example, by proteolytic cleavage, or by
chemical linking. For example, an antigen-binding fragment (e.g.
Fab') of an antibody may be prepared and converted to Fab'-thiol
derivative and then mixed and reacted with another converted Fab'
derivative having a different antigenic specificity to form a
bispecific antibody molecule (see, for example, Brennan et al.,
Science, 229: 81 (1985)).
[0512] In certain embodiments, the bispecific antibody molecules
may be engineered to promote heavy chain heterodimerization of the
two different antigen-binding sites. In certain embodiments, the Fc
region is modified at the interface so that a knob-into-hole
association can be formed to promote heterodimerization.
"Knob-into-hole" as used herein, refers to an interaction between
two polypeptides (such as CH3 domain), where one polypeptide has a
protuberance (i.e. "knob") due to presence of an amino acid residue
having a bulky side chain (e.g. tyrosine or tryptophan), and the
other polypeptide has a cavity (i.e. "hole") where a small side
chain amino acid residue resides (e.g. alanine or threonine), and
the protuberance is positionable in the cavity so as to promote
interaction of the two polypeptides to form a heterodimer or a
complex. Methods of generating polypeptides with knobs-into-holes
are known in the art, e.g., as described in U.S. Pat. No.
5,731,168.
[0513] In some embodiments, "charged pairs" can be introduced to
the Fc polypeptides to electrostatically steer the formation
towards heterodimerization. Exemplary pairs include,
D221E/P228E/L368E paired with D221R/P228R/K409R and
C220E/P228E/368E paired with C220R/E224R/P228R/K409R (see
Gunasekaran et al., 2010, J. Biol. Chem. 285(25):19637.).
[0514] In some embodiments, the binding interface of the two Fc
polypeptide chains can be engineered such that in the heterodimer
configuration, residues interact with residues of similar physical
property (e.g., polar residues interacting with polar residues, or
hydrophobic residues interact with hydrophobic residues), while in
the homodimer configuration residues interact with residues of
different physical property. Exemplary modifications include
substitution at positions 364, 368, 399, 405, 409, 411, or any
combination thereof (see, e.g, WO2014/145806, WO2014/110601,
WO2016/086186, WO2016/086189, WO2016/086196, and
WO2016/182751).
[0515] In some embodiments, the bispecific antibody molecules may
be engineered to reduce random pairing of two different light chain
variable regions with the two different heavy chain variable
regions. In some embodiments, the bispecific antibody molecule
comprise a common light chain capable of pairing with the two heavy
chain variable regions. In some other embodiments, CH1 domain of
one heavy chain is exchanged with the constant region (CL) of the
corresponding light chain (such as that applied in CrossMab
technology). In some other embodiments, mutations are introduced
into the CH1-CL interface and/or the VH-VL interface of the Fab
fragments, so as to enforce correct pairing of the light chains
with the corresponding heavy chains. In some other embodiments, the
CH1 domain and CL domain in one antigen-binding domain are replaced
by TCR constant domains, so as to minimize mispairing between heavy
chain of the first antigen-binding domain and light chain of the
second antigen-binding domain (such as that applied in WuxiBody
technology).
[0516] In some embodiments, the modified antibody or an
antigen-binding fragment thereof of this disclosure, wherein the
antigen-binding domain can comprise: [0517] a. 1, 2, or 3 heavy
chain CDR sequences selected from SEQ ID NO: 1, SEQ ID NO: 2, and
SEQ ID NO: 3; [0518] b. 1, 2, or 3 heavy chain CDR sequences
selected from SEQ ID NO: 11, SEQ ID NO: 12, and SEQ ID NO: 13;
[0519] c. 1, 2, or 3 heavy chain CDR sequences selected from SEQ ID
NO: 21, SEQ ID NO: 22, and SEQ ID NO: 23; [0520] d. 1, 2, or 3
heavy chain CDR sequences selected from SEQ ID NO: 31, SEQ ID NO:
32, and SEQ ID NO: 33; [0521] e. 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 41, SEQ ID NO: 42, and SEQ ID
NO: 43; [0522] f. 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 51, SEQ ID NO: 52, and SEQ ID NO: 53; [0523] g. 1,
2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 65, SEQ
ID NO: 66, and SEQ ID NO: 67; [0524] h. 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 75, SEQ ID NO: 76, and SEQ ID
NO: 77; [0525] i. 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 85, SEQ ID NO: 86, and SEQ ID NO: 87; [0526] j. 1,
2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 95, SEQ
ID NO: 96, and SEQ ID NO: 97; [0527] k. 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 105, SEQ ID NO: 106, and SEQ ID
NO: 107; [0528] l. 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 136, SEQ ID NO: 137, and SEQ ID NO: 138; [0529] m.
1, 2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 146,
SEQ ID NO: 147, and SEQ ID NO: 148; [0530] n. 1, 2, or 3 heavy
chain CDR sequences selected from SEQ ID NO: 156, SEQ ID NO: 157,
and SEQ ID NO: 158; [0531] o. 1, 2, or 3 heavy chain CDR sequences
selected from SEQ ID NO: 166, SEQ ID NO: 167, and SEQ ID NO: 168;
[0532] p. 1, 2, or 3 heavy chain CDR sequences selected from SEQ ID
NO: 176, SEQ ID NO: 177, and SEQ ID NO: 178; [0533] q. 1, 2, or 3
heavy chain CDR sequences selected from SEQ ID NO: 186, SEQ ID NO:
187, and SEQ ID NO: 188; [0534] r. 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 196, SEQ ID NO: 197, and SEQ ID
NO: 198; [0535] s. 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 206, SEQ ID NO: 207, and SEQ ID NO: 208; [0536] t.
1, 2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 216,
SEQ ID NO: 217, and SEQ ID NO: 218; [0537] u. 1, 2, or 3 heavy
chain CDR sequences selected from SEQ ID NO: 226, SEQ ID NO: 227,
and SEQ ID NO: 228; [0538] v. 1, 2, or 3 heavy chain CDR sequences
selected from SEQ ID NO: 236, SEQ ID NO: 237, and SEQ ID NO: 238;
[0539] w. 1, 2, or 3 heavy chain CDR sequences selected from SEQ ID
NO: 246, SEQ ID NO: 247, and SEQ ID NO: 248; [0540] x. 1, 2, or 3
heavy chain CDR sequences selected from SEQ ID NO: 256, SEQ ID NO:
257, and SEQ ID NO: 258; [0541] y. 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 266, SEQ ID NO: 267, and SEQ ID
NO: 268; [0542] z. 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 276, SEQ ID NO: 277, and SEQ ID NO: 278; [0543] aa.
1, 2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 286,
SEQ ID NO: 287, and SEQ ID NO: 288; [0544] bb. 1, 2, or 3 heavy
chain CDR sequences selected from SEQ ID NO: 296, SEQ ID NO: 297,
and SEQ ID NO: 298; [0545] cc. 1, 2, or 3 heavy chain CDR sequences
selected from SEQ ID NO: 306, SEQ ID NO: 307, and SEQ ID NO: 308;
[0546] dd. 1, 2, or 3 heavy chain CDR sequences selected from SEQ
ID NO: 316, SEQ ID NO: 317, and SEQ ID NO: 318; [0547] ee. 1, 2, or
3 heavy chain CDR sequences selected from SEQ ID NO: 326, SEQ ID
NO: 327, and SEQ ID NO: 328; [0548] . 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 336, SEQ ID NO: 337, and SEQ ID
NO: 338; [0549] gg. 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 346, SEQ ID NO: 347, and SEQ ID NO: 348; [0550] hh.
1, 2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 356,
SEQ ID NO: 357, and SEQ ID NO: 358; [0551] ii. 1, 2, or 3 heavy
chain CDR sequences selected from SEQ ID NO: 366, SEQ ID NO: 367,
and SEQ ID NO: 368; [0552] jj. 1, 2, or 3 heavy chain CDR sequences
selected from SEQ ID NO: 376, SEQ ID NO: 377, and SEQ ID NO: 378;
[0553] kk. 1, 2, or 3 heavy chain CDR sequences selected from SEQ
ID NO: 386, SEQ ID NO: 387, and SEQ ID NO: 388; [0554] ll. 1, 2, or
3 heavy chain CDR sequences selected from SEQ ID NO: 396, SEQ ID
NO: 397, and SEQ ID NO: 398; [0555] mm. 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 406, SEQ ID NO: 407, and SEQ ID
NO: 408; [0556] nn. 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 416, SEQ ID NO: 417, and SEQ ID NO: 418; [0557] oo.
1, 2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 426,
SEQ ID NO: 427, and SEQ ID NO: 428; or
[0558] a combination thereof.
[0559] In some embodiments, the modified antibody or antigen
binding fragment disclosed above, wherein the antigen-binding
domain comprises: [0560] a. 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6; [0561]
b. 1, 2, or 3 light chain CDR sequences selected from SEQ ID NO:
14, SEQ ID NO: 15, and SEQ ID NO: 16; [0562] c. 1, 2, or 3 light
chain CDR sequences selected from SEQ ID NO: 24, SEQ ID NO: 25 and
SEQ ID NO: 26; [0563] d. 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36;
[0564] e. 1, 2, or 3 light chain CDR sequences selected from SEQ ID
NO: 44, SEQ ID NO: 45 and SEQ ID NO: 46; [0565] f. 1, 2, or 3 light
chain CDR sequences selected from SEQ ID NO: 54, SEQ ID NO: 55 and
SEQ ID NO: 56; [0566] g. 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 68, SEQ ID NO: 69, and SEQ ID NO: 70;
[0567] h. 1, 2, or 3 light chain CDR sequences selected from SEQ ID
NO: 78, SEQ ID NO: 79, and SEQ ID NO: 80; [0568] i. 1, 2, or 3
light chain CDR sequences selected from SEQ ID NO: 88, SEQ ID NO:
89, and SEQ ID NO: 90. [0569] j. 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 98, SEQ ID NO: 99, and SEQ ID
NO: 100; [0570] k. 1, 2, or 3 light chain CDR sequences selected
from SEQ ID NO: 108, SEQ ID NO: 109, and SEQ ID NO: 110; [0571] l.
1, 2, or 3 light chain CDR sequences selected from SEQ ID NO: 139,
SEQ ID NO: 140, and SEQ ID NO: 141; [0572] m. 1, 2, or 3 light
chain CDR sequences selected from SEQ ID NO: 149, SEQ ID NO: 150,
and SEQ ID NO: 151; [0573] n. 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 159, SEQ ID NO: 160, and SEQ ID NO: 161;
[0574] o. 1, 2, or 3 light chain CDR sequences selected from SEQ ID
NO: 169, SEQ ID NO: 170, and SEQ ID NO: 171; [0575] p. 1, 2, or 3
light chain CDR sequences selected from SEQ ID NO: 179, SEQ ID NO:
180, and SEQ ID NO: 181; [0576] q. 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 189, SEQ ID NO: 190, and SEQ ID
NO: 191; [0577] r. 1, 2, or 3 light chain CDR sequences selected
from SEQ ID NO: 199, SEQ ID NO: 200, and SEQ ID NO: 201; [0578] s.
1, 2, or 3 light chain CDR sequences selected from SEQ ID NO: 209,
SEQ ID NO: 210, and SEQ ID NO: 211; [0579] t. 1, 2, or 3 light
chain CDR sequences selected from SEQ ID NO: 219, SEQ ID NO: 220,
and SEQ ID NO: 221; [0580] u. 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 229, SEQ ID NO: 230, and SEQ ID NO: 231;
[0581] v. 1, 2, or 3 light chain CDR sequences selected from SEQ ID
NO: 239, SEQ ID NO: 240, and SEQ ID NO: 241; [0582] w. 1, 2, or 3
light chain CDR sequences selected from SEQ ID NO: 249, SEQ ID NO:
250, and SEQ ID NO: 251; [0583] x. 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 259, SEQ ID NO: 260, and SEQ ID
NO: 261; [0584] y. 1, 2, or 3 light chain CDR sequences selected
from SEQ ID NO: 269, SEQ ID NO: 270, and SEQ ID NO: 271; [0585] z.
1, 2, or 3 light chain CDR sequences selected from SEQ ID NO: 279,
SEQ ID NO: 280, and SEQ ID NO: 281; [0586] aa. 1, 2, or 3 light
chain CDR sequences selected from SEQ ID NO: 289, SEQ ID NO: 290,
and SEQ ID NO: 291; [0587] bb. 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 299, SEQ ID NO: 300, and SEQ ID NO: 301;
[0588] cc. 1, 2, or 3 light chain CDR sequences selected from SEQ
ID NO: 309, SEQ ID NO: 310, and SEQ ID NO: 311; [0589] dd. 1, 2, or
3 light chain CDR sequences selected from SEQ ID NO: 319, SEQ ID
NO: 320, and SEQ ID NO: 321; [0590] ee. 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 329, SEQ ID NO: 330, and SEQ ID
NO: 331; [0591] ff. 1, 2, or 3 light chain CDR sequences selected
from SEQ ID NO: 339, SEQ ID NO: 340, and SEQ ID NO: 341; [0592] gg.
1, 2, or 3 light chain CDR sequences selected from SEQ ID NO: 349,
SEQ ID NO: 350, and SEQ ID NO: 351; [0593] hh. 1, 2, or 3 light
chain CDR sequences selected from SEQ ID NO: 359, SEQ ID NO: 360,
and SEQ ID NO: 361; [0594] ii. 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 369, SEQ ID NO: 370, and SEQ ID NO: 371;
[0595] jj. 1, 2, or 3 light chain CDR sequences selected from SEQ
ID NO: 379, SEQ ID NO: 380, and SEQ ID NO: 381; [0596] kk. 1, 2, or
3 light chain CDR sequences selected from SEQ ID NO: 389, SEQ ID
NO: 390, and SEQ ID NO: 391; [0597] ll. 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 399, SEQ ID NO: 400, and SEQ ID
NO: 401; [0598] mm. 1, 2, or 3 light chain CDR sequences selected
from SEQ ID NO: 409, SEQ ID NO: 410, and SEQ ID NO: 411; [0599] nn.
1, 2, or 3 light chain CDR sequences selected from SEQ ID NO: 419,
SEQ ID NO: 420, and SEQ ID NO: 421; [0600] oo. 1, 2, or 3 light
chain CDR sequences selected from SEQ ID NO: 429, SEQ ID NO: 430,
and SEQ ID NO: 431; or
[0601] a combination thereof.
[0602] In some embodiments, the modified antibody or an
antigen-binding fragment thereof disclosed herein, wherein the
antigen-binding domain can comprise: [0603] a. a heavy chain CDR1
(HCDR1) comprising the sequence of SEQ ID NO: 1, a heavy chain CDR2
(HCDR2) comprising the sequence of SEQ ID NO: 2, a heavy chain CDR3
(HCDR3) comprising the sequence of SEQ ID NO: 3; a light chain CDR1
(LCDR1) comprising the sequence of SEQ ID NO: 4, a light chain CDR2
(LCDR2) comprising the sequence of SEQ ID NO: 5, and a light chain
CDR3 (LCDR3) comprising the sequence of SEQ ID NO: 6; [0604] b. a
HCDR1 comprising the sequence of SEQ ID NO: 11, a HCDR2 comprising
the sequence of SEQ ID NO: 12, a HCDR3 comprising the sequence of
SEQ ID NO: 13, a LCDR1 comprising the sequence of SEQ ID NO: 14, a
LCDR2 comprising the sequence of SEQ ID NO: 15, and a LCDR3
comprising the sequence of SEQ ID NO: 16; [0605] c. a HCDR1
comprising the sequence of SEQ ID NO: 21, a HCDR2 comprising the
sequence of SEQ ID NO: 22, a HCDR3 comprising the sequence of SEQ
ID NO: 23, a LCDR1 comprising the sequence of SEQ ID NO: 24, a
LCDR2 comprising the sequence of SEQ ID NO: 25, and a LCDR3
comprising the sequence of SEQ ID NO: 26; [0606] d. a HCDR1
comprising the sequence of SEQ ID NO: 31, a HCDR2 comprising the
sequence of SEQ ID NO: 32, a HCDR3 comprising the sequence of SEQ
ID NO: 33, a LCDR1 comprising the sequence of SEQ ID NO: 34, a
LCDR2 comprising the sequence of SEQ ID NO: 35, and a LCDR3
comprising the sequence of SEQ ID NO: 36; [0607] e. a HCDR1
comprising the sequence of SEQ ID NO: 41, a HCDR2 comprising the
sequence of SEQ ID NO: 42, a HCDR3 comprising the sequence of SEQ
ID NO: 43, a LCDR1 comprising the sequence of SEQ ID NO: 44, a
LCDR2 comprising the sequence of SEQ ID NO: 45, and a LCDR3
comprising the sequence of SEQ ID NO: 46; [0608] f. a HCDR1
comprising the sequence of SEQ ID NO: 51, a HCDR2 comprising the
sequence of SEQ ID NO: 52, a HCDR3 comprising the sequence of SEQ
ID NO: 53, a LCDR1 comprising the sequence of SEQ ID NO: 54, a
LCDR2 comprising the sequence of SEQ ID NO: 55, and a LCDR3
comprising the sequence of SEQ ID NO: 56; [0609] g. a HCDR1
comprising the sequence of SEQ ID NO: 65, a HCDR2 comprising the
sequence of SEQ ID NO: 66, a HCDR3 comprising the sequence of SEQ
ID NO: 67, a LCDR1 comprising the sequence of SEQ ID NO: 68, a
LCDR2 comprising the sequence of SEQ ID NO: 69, and a LCDR3
comprising the sequence of SEQ ID NO: 70; [0610] h. a HCDR1
comprising the sequence of SEQ ID NO: 75, a HCDR2 comprising the
sequence of SEQ ID NO: 76, a HCDR3 comprising the sequence of SEQ
ID NO: 77, a LCDR1 comprising the sequence of SEQ ID NO: 78, a
LCDR2 comprising the sequence of SEQ ID NO: 79, and a LCDR3
comprising the sequence of SEQ ID NO: 80; [0611] i. a HCDR1
comprising the sequence of SEQ ID NO: 85, a HCDR2 comprising the
sequence of SEQ ID NO: 86, a HCDR3 comprising the sequence of SEQ
ID NO: 87, a LCDR1 comprising the sequence of SEQ ID NO: 88, a
LCDR2 comprising the sequence of SEQ ID NO: 89, and a LCDR3
comprising the sequence of SEQ ID NO: 90; [0612] j. a HCDR1
comprising the sequence of SEQ ID NO: 95, a HCDR2 comprising the
sequence of SEQ ID NO: 96, a HCDR3 comprising the sequence of SEQ
ID NO: 97, a LCDR1 comprising the sequence of SEQ ID NO: 98, a
LCDR2 comprising the sequence of SEQ ID NO: 99, and a LCDR3
comprising the sequence of SEQ ID NO: 100; [0613] k. a HCDR1
comprising the sequence of SEQ ID NO: 105, a HCDR2 comprising the
sequence of SEQ ID NO: 106, a HCDR3 comprising the sequence of SEQ
ID NO: 107, a LCDR1 comprising the sequence of SEQ ID NO: 108, a
LCDR2 comprising the sequence of SEQ ID NO: 109, and a LCDR3
comprising the sequence of SEQ ID NO: 110; [0614] l. a HCDR1
comprising the sequence of SEQ ID NO: 136, a HCDR2 comprising the
sequence of SEQ ID NO: 137, a HCDR3 comprising the sequence of SEQ
ID NO: 138, a LCDR1 comprising the sequence of SEQ ID NO: 139, a
LCDR2 comprising the sequence of SEQ ID NO: 140, and a LCDR3
comprising the sequence of SEQ ID NO: 141; [0615] m. HCDR1
comprising the sequence of SEQ ID NO: 146, a HCDR2 comprising the
sequence of SEQ ID NO: 147, a HCDR3 comprising the sequence of SEQ
ID NO: 148, a LCDR1 comprising the sequence of SEQ ID NO: 149, a
LCDR2 comprising the sequence of SEQ ID NO: 150, and a LCDR3
comprising the sequence of SEQ ID NO: 151; [0616] n. HCDR1
comprising the sequence of SEQ ID NO: 156, a HCDR2 comprising the
sequence of SEQ ID NO: 157, a HCDR3 comprising the sequence of SEQ
ID NO: 158, a LCDR1 comprising the sequence of SEQ ID NO: 159, a
LCDR2 comprising the sequence of SEQ ID NO: 160, and a LCDR3
comprising the sequence of SEQ ID NO: 161; [0617] o. HCDR1
comprising the sequence of SEQ ID NO: 166, a HCDR2 comprising the
sequence of SEQ ID NO: 167, a HCDR3 comprising the sequence of SEQ
ID NO: 168, a LCDR1 comprising the sequence of SEQ ID NO: 169, a
LCDR2 comprising the sequence of SEQ ID NO: 170, and a LCDR3
comprising the sequence of SEQ ID NO: 171; [0618] p. HCDR1
comprising the sequence of SEQ ID NO: 176, a HCDR2 comprising the
sequence of SEQ ID NO: 177, a HCDR3 comprising the sequence of SEQ
ID NO: 178, a LCDR1 comprising the sequence of SEQ ID NO: 179, a
LCDR2 comprising the sequence of SEQ ID NO: 180, and a LCDR3
comprising the sequence of SEQ ID NO: 181; [0619] q. HCDR1
comprising the sequence of SEQ ID NO: 186, a HCDR2 comprising the
sequence of SEQ ID NO: 187, a HCDR3 comprising the sequence of SEQ
ID NO: 188, a LCDR1 comprising the sequence of SEQ ID NO: 189, a
LCDR2 comprising the sequence of SEQ ID NO: 190, and a LCDR3
comprising the sequence of SEQ ID NO: 191; [0620] r. HCDR1
comprising the sequence of SEQ ID NO: 196, a HCDR2 comprising the
sequence of SEQ ID NO: 197, a HCDR3 comprising the sequence of SEQ
ID NO: 198, a LCDR1 comprising the sequence of SEQ ID NO: 199, a
LCDR2 comprising the sequence of SEQ ID NO: 200, and a LCDR3
comprising the sequence of SEQ ID NO: 201; [0621] s. HCDR1
comprising the sequence of SEQ ID NO: 206, a HCDR2 comprising the
sequence of SEQ ID NO: 207, a HCDR3 comprising the sequence of SEQ
ID NO: 208, a LCDR1 comprising the sequence of SEQ ID NO: 209, a
LCDR2 comprising the sequence of SEQ ID NO: 210, and a LCDR3
comprising the sequence of SEQ ID NO: 211; [0622] t. HCDR1
comprising the sequence of SEQ ID NO: 216, a HCDR2 comprising the
sequence of SEQ ID NO: 217, a HCDR3 comprising the sequence of SEQ
ID NO: 218, a LCDR1 comprising the sequence of SEQ ID NO: 219, a
LCDR2 comprising the sequence of SEQ ID NO: 220, and a LCDR3
comprising the sequence of SEQ ID NO: 221; [0623] u. HCDR1
comprising the sequence of SEQ ID NO: 226, a HCDR2 comprising the
sequence of SEQ ID NO: 227, a HCDR3 comprising the sequence of SEQ
ID NO: 228, a LCDR1 comprising the sequence of SEQ ID NO: 229, a
LCDR2 comprising the sequence of SEQ ID NO: 230, and a LCDR3
comprising the sequence of SEQ ID NO: 231; [0624] v. HCDR1
comprising the sequence of SEQ ID NO: 236, a HCDR2 comprising the
sequence of SEQ ID NO: 237, a HCDR3 comprising the sequence of SEQ
ID NO: 238, a LCDR1 comprising the sequence of SEQ ID NO: 239, a
LCDR2 comprising the sequence of SEQ ID NO: 240, and a LCDR3
comprising the sequence of SEQ ID NO: 241; [0625] w. HCDR1
comprising the sequence of SEQ ID NO: 246, a HCDR2 comprising the
sequence of SEQ ID NO: 247, a HCDR3 comprising the sequence of SEQ
ID NO: 248, a LCDR1 comprising the sequence of SEQ ID NO: 249, a
LCDR2 comprising the sequence of SEQ ID NO: 250, and a LCDR3
comprising the sequence of SEQ ID NO: 251; [0626] x. HCDR1
comprising the sequence of SEQ ID NO: 256, a HCDR2 comprising the
sequence of SEQ ID NO: 257, a HCDR3 comprising the sequence of SEQ
ID NO: 258, a LCDR1 comprising the sequence of SEQ ID NO: 259, a
LCDR2 comprising the sequence of SEQ ID NO: 260, and a LCDR3
comprising the sequence of SEQ ID NO: 261; [0627] y. HCDR1
comprising the sequence of SEQ ID NO: 266, a HCDR2 comprising the
sequence of SEQ ID NO: 267, a HCDR3 comprising the sequence of SEQ
ID NO: 268, a LCDR1 comprising the sequence of SEQ ID NO: 269, a
LCDR2 comprising the sequence of SEQ ID NO: 270, and a LCDR3
comprising the sequence of SEQ ID NO: 271; [0628] z. HCDR1
comprising the sequence of SEQ ID NO: 276, a HCDR2 comprising the
sequence of SEQ ID NO: 277, a HCDR3 comprising the sequence of SEQ
ID NO: 278, a LCDR1 comprising the sequence of SEQ ID NO: 279, a
LCDR2 comprising the sequence of SEQ ID NO: 280, a LCDR3 comprising
the sequence of SEQ ID NO: 281; [0629] aa. HCDR1 comprising the
sequence of SEQ ID NO: 286, a HCDR2 comprising the sequence of SEQ
ID NO: 287, a HCDR3 comprising the sequence of SEQ ID NO: 288, a
LCDR1 comprising the sequence of SEQ ID NO: 289, a LCDR2 comprising
the sequence of SEQ ID NO: 290, a LCDR3 comprising the sequence of
SEQ ID NO: 291; [0630] bb. HCDR1 comprising the sequence of SEQ ID
NO: 296, a HCDR2 comprising the sequence of SEQ ID NO: 297, a HCDR3
comprising the sequence of SEQ ID NO: 298, a LCDR1 comprising the
sequence of SEQ ID NO: 299, a LCDR2 comprising the sequence of SEQ
ID NO: 300, a LCDR3 comprising the sequence of SEQ ID NO: 301;
[0631] cc. HCDR1 comprising the sequence of SEQ ID NO: 306, a HCDR2
comprising the sequence of SEQ ID NO: 307, a HCDR3 comprising the
sequence of SEQ ID NO: 308, a LCDR1 comprising the sequence of SEQ
ID NO: 309, a LCDR2 comprising the sequence of SEQ ID NO: 310, a
LCDR3 comprising the sequence of SEQ ID NO: 311; [0632] dd. HCDR1
comprising the sequence of SEQ ID NO: 316, a HCDR2 comprising the
sequence of SEQ ID NO: 317, a HCDR3 comprising the sequence of SEQ
ID NO: 318, a LCDR1 comprising the sequence of SEQ ID NO: 319, a
LCDR2 comprising the sequence of SEQ ID NO: 320, a LCDR3 comprising
the sequence of SEQ ID NO: 321; [0633] ee. HCDR1 comprising the
sequence of SEQ ID NO: 326, a HCDR2 comprising the sequence of SEQ
ID NO: 327, a HCDR3 comprising the sequence of SEQ ID NO: 328, a
LCDR1 comprising the sequence of SEQ ID NO: 329, a LCDR2 comprising
the sequence of SEQ ID NO: 330, a LCDR3 comprising the sequence of
SEQ ID NO: 331; [0634] ff. HCDR1 comprising the sequence of SEQ ID
NO: 336, a HCDR2 comprising the sequence of SEQ ID NO: 337, a HCDR3
comprising the sequence of SEQ ID NO: 338, a LCDR1 comprising the
sequence of SEQ ID NO: 339, a LCDR2 comprising the sequence of SEQ
ID NO: 340, a LCDR3 comprising the sequence of SEQ ID NO: 341;
[0635] gg. HCDR1 comprising the sequence of SEQ ID NO: 346, a HCDR2
comprising the sequence of SEQ ID NO: 347, a HCDR3 comprising the
sequence of SEQ ID NO: 348, a LCDR1 comprising the sequence of SEQ
ID NO: 349, a LCDR2 comprising the sequence of SEQ ID NO: 350, a
LCDR3 comprising the sequence of SEQ ID NO: 351; [0636] hh. HCDR1
comprising the sequence of SEQ ID NO: 356, a HCDR2 comprising the
sequence of SEQ ID NO: 357, a HCDR3 comprising the sequence of SEQ
ID NO: 358, a LCDR1 comprising the sequence of SEQ ID NO: 359, a
LCDR2 comprising the sequence of SEQ ID NO: 360, a LCDR3 comprising
the sequence of SEQ ID NO: 361; [0637] ii. HCDR1 comprising the
sequence of SEQ ID NO: 366, a HCDR2 comprising the sequence of SEQ
ID NO: 367, a HCDR3 comprising the sequence of SEQ ID NO: 368, a
LCDR1 comprising the sequence of SEQ ID NO: 369, a LCDR2 comprising
the sequence of SEQ ID NO: 370, a LCDR3 comprising the sequence of
SEQ ID NO: 371; [0638] jj. HCDR1 comprising the sequence of SEQ ID
NO: 376, a HCDR2 comprising the sequence of SEQ ID NO: 377, a HCDR3
comprising the sequence of SEQ ID NO: 378, a LCDR1 comprising the
sequence of SEQ ID NO: 379, a LCDR2 comprising the sequence of SEQ
ID NO: 380, a LCDR3 comprising the sequence of SEQ ID NO: 381;
[0639] kk. HCDR1 comprising the sequence of SEQ ID NO: 386, a HCDR2
comprising the sequence of SEQ ID NO: 387, a HCDR3 comprising the
sequence of SEQ ID NO: 388, a LCDR1 comprising the sequence of SEQ
ID NO: 389, a LCDR2 comprising the sequence of SEQ ID NO: 390, a
LCDR3 comprising the sequence of SEQ ID NO: 391; [0640] ll. HCDR1
comprising the sequence of SEQ ID NO: 396, a HCDR2 comprising the
sequence of SEQ ID NO: 397, a HCDR3 comprising the sequence of SEQ
ID NO: 398, a LCDR1 comprising the sequence of SEQ ID NO: 399, a
LCDR2 comprising the sequence of SEQ ID NO: 400, a LCDR3 comprising
the sequence of SEQ ID NO: 401; [0641] mm. HCDR1 comprising the
sequence of SEQ ID NO: 406, a HCDR2 comprising the sequence of SEQ
ID NO: 407, a HCDR3 comprising the sequence of SEQ ID NO: 408, a
LCDR1 comprising the sequence of SEQ ID NO: 409, a LCDR2 comprising
the sequence of SEQ ID NO: 410, a LCDR3 comprising the sequence of
SEQ ID NO: 411; [0642] nn. HCDR1 comprising the sequence of SEQ ID
NO: 416, a HCDR2 comprising the sequence of SEQ ID NO: 417, a HCDR3
comprising the sequence of SEQ ID NO: 418, a LCDR1 comprising the
sequence of SEQ ID NO: 419, a LCDR2 comprising the sequence of SEQ
ID NO: 420, a LCDR3 comprising the sequence of SEQ ID NO: 421;
[0643] oo. HCDR1 comprising the sequence of SEQ ID NO: 426, a HCDR2
comprising the sequence of SEQ ID NO: 427, a HCDR3 comprising the
sequence of SEQ ID NO: 428, a LCDR1 comprising the sequence of SEQ
ID NO: 429, a LCDR2 comprising the sequence of SEQ ID NO: 430, a
LCDR3 comprising the sequence of SEQ ID NO: 431; or
[0644] a combination thereof.
[0645] In some embodiments, the modified antibody or an
antigen-binding fragment thereof disclosed herein, wherein the
antigen-binding domain can comprise:
[0646] a HCDR1 comprising the sequence of SEQ ID NO: 105, a HCDR2
comprising the sequence of SEQ ID NO: 106, a HCDR3 comprising the
sequence of SEQ ID NO: 107, a LCDR1 comprising the sequence of SEQ
ID NO: 108, a LCDR2 comprising the sequence of SEQ ID NO: 109, and
a LCDR3 comprising the sequence of SEQ ID NO: 110;
[0647] a HCDR1 comprising the sequence of SEQ ID NO: 136, a HCDR2
comprising the sequence of SEQ ID NO: 137, a HCDR3 comprising the
sequence of SEQ ID NO: 138, a LCDR1 comprising the sequence of SEQ
ID NO: 139, a LCDR2 comprising the sequence of SEQ ID NO: 140, and
a LCDR3 comprising the sequence of SEQ ID NO: 141;
[0648] or a combination thereof.
[0649] In some embodiments, the modified antibody can comprise a
first antigen-binding domain comprising a HCDR1 comprising the
sequence of SEQ ID NO: 105, a HCDR2 comprising the sequence of SEQ
ID NO: 106, a HCDR3 comprising the sequence of SEQ ID NO: 107, a
LCDR1 comprising the sequence of SEQ ID NO: 108, a LCDR2 comprising
the sequence of SEQ ID NO: 109, and a LCDR3 comprising the sequence
of SEQ ID NO: 110; and a second antigen-binding domain comprising a
HCDR1 comprising the sequence of SEQ ID NO: 136, a HCDR2 comprising
the sequence of SEQ ID NO: 137, a HCDR3 comprising the sequence of
SEQ ID NO: 138, a LCDR1 comprising the sequence of SEQ ID NO: 139,
a LCDR2 comprising the sequence of SEQ ID NO: 140, and a LCDR3
comprising the sequence of SEQ ID NO: 141; and wherein the antibody
comprises the modified human IgG constant domain comprises a
substitution with tyrosine at amino acid residue 252, a
substitution with threonine at amino acid residue 254, and a
substitution with glutamic acid at amino acid residue 256, numbered
according to the EU index as in Kabat.
[0650] In some embodiments, the modified antibody or an
antigen-binding fragment thereof disclosed herein, wherein the
modified antibody or the antigen-binding fragment can have a
half-life (T.sub.1/2) in a range of from 50 to 120 days in vivo,
such as in a human subject. The modified antibody or the
antigen-binding fragment can have a half-life (T.sub.1/2) in a
range of from 50 to 120 days, 60 to 120 days, 70 to 120 days, 80 to
120 days, 90 to 120 days, 100 to 120 day, or 110 to 120 days.
[0651] In some embodiments, the modified antibody or an
antigen-binding fragment thereof disclosed herein, wherein the
modified antibody can comprise at least one amino acid subsequent
substitutions in the antigen-binding domain, the human IgG constant
domain, a light chain of the modified antibody, a heavy chain of
the modified antibody, or a combination thereof. In some cases, the
subsequent substitution can comprise substituting a cystine residue
to a non-cystine residue. In some cases, the cystine residue can be
substituted with a serine residue. In some embodiments, a modified
antibody can comprise a C106S substitution, wherein the cystine 106
is substituted with a serine, in the heavy chain variable region
HDR3, numbered according to the international ImMunoGeneTics system
(IMGT) unique numbering.
[0652] In some embodiments, a modified antibody can comprise an
antigen-binding domain having an antigen-binding affinity and a
covalently linked modified human IgG constant domain, wherein the
antigen-binding affinity comprises at least one of the LCDRs and at
least one of the HCDRs listed in Table 1, wherein the modified
human IgG constant domain comprises a substitution with tyrosine at
amino acid residue 252, a substitution with threonine at amino acid
residue 254, and a substitution with glutamic acid at amino acid
residue 256, numbered according to the EU index as in Kabat, and a
C106S substitution, wherein the cystine 106 is substituted with a
serine, in the heavy chain variable region HDR3, numbered according
to the international ImMunoGeneTics system (IMGT) unique numbering.
In some embodiments, a modified antibody can comprise an
antigen-binding domain having an antigen-binding affinity and a
covalently linked modified human IgG constant domain, wherein the
antigen-binding affinity comprises HCDR SEQ ID No.: 136, SEQ ID
No.: 137, SEQ ID No.: 138, LCDR SEQ ID No.: 139, SEQ ID. No.: 140
and SEQ ID. No.: 141 (P2B-1G5), wherein the modified human IgG
constant domain comprises a substitution with tyrosine at amino
acid residue 252, a substitution with threonine at amino acid
residue 254, and a substitution with glutamic acid at amino acid
residue 256, numbered according to the EU index as in Kabat, and a
C106S substitution, wherein the cystine 106 is substituted with a
serine, in the heavy chain variable region, numbered according to
the international ImMunoGeneTics system (IMGT) unique
numbering.
[0653] In some embodiments, the modified antibody or an
antigen-binding fragment thereof disclosed herein can further
comprise one or more subsequent modified antibodies selected from a
first subsequent modified antibody comprising two antigen-binding
domains each having a same or different affinity to the SARS-CoV-2,
a second subsequent modified antibody comprising a first
antigen-binding domain having a binding affinity to the SARS-CoV-2
and a second antigen-binding domain having a binding affinity to a
second pathogen that is different from the SARS-CoV-2, a third
subsequent modified antibody comprising two antigen-binding domains
each having a binding affinity to the second pathogen, or a
combination thereof. The term "different affinities to the
SARS-CoV-2" refers affinity that can bind to a different epitope or
binding site of the SARS-CoV-2, a different affinity level that can
bind to the same epitope or binding site of the SARS-CoV-2, or a
combination thereof.
[0654] In some embodiments, the modified antibody or an
antigen-binding fragment thereof disclosed herein, wherein the
binding affinity to the second pathogen can be selected from a
binding affinity to SARS-CoV, MERS-CoV, one or more bacteria, one
or more fungus, one or more viruses, one or more parasites, a part
thereof, or a combination thereof.
[0655] In some embodiments, the modified antibody or an
antigen-binding fragment thereof disclosed herein, wherein the
modified antibody or the antigen-binding fragment thereof can be a
single chain antibody, a diabody, a Fab, a Fab', a F(ab')2, a Fd,
an Fv fragment, a disulfide stabilized Fv fragment (dsFv), a
(dsFv)2, a bispecific dsFv (dsFv-dsFv'), a disulfide stabilized
diabody (ds diabody), a single-chain antibody molecule (scFv), an
scFv dimer (bivalent diabody), a bispecific scFv dimer, a
multispecific antibody, a heavy chain antibody, a camelized single
domain antibody, a nanobody, a domain antibody, or a bivalent
domain antibody, as disclosed above and hereafter.
[0656] Competitive Binding, Crystal Structure and Epitope
[0657] In one aspect, the present disclosure provides an isolated
or recombinant antibody or an antigen-binding fragment thereof,
which competes for binding to RBD of spike protein of SARS-CoV-2
with the antibody or an antigen-binding fragment thereof described
herein.
[0658] Antibodies or antigen binding fragments that competes with
the antibody or antigen-binding fragment provided herein for
binding to RBD of spike protein of SARS-CoV-2 include, but are not
limited to, antibodies, antibody fragments and other binding agents
that bind to an epitope or binding site bound by the antibody or
antigen-binding fragment provided herein, or bind to a sufficiently
proximal epitope or binding site. Preferably, competitive
antibodies or antigen binding fragments of the disclosure will,
when present in excess, inhibit specific binding of the antibody or
antigen-binding fragment provided herein to RBD of the spike
protein of SARS-CoV-2 by at least 10%, preferably by at least 25%,
more preferably by at least 50%, and most preferably by at least
75%-90% or even greater. The identification of one or more
competitive antibodies or antigen binding fragments that bind to
about, substantially, essentially or at the same epitope as the
antibodies or antigen binding fragments of the present disclosure
is a straightforward technical matter. As the identification of
competitive binding molecules is determined in comparison to a
reference binding molecule, for example, the antibodies or antigen
binding fragments of the present disclosure, it will be understood
that actually determining the epitope to which the reference
binding molecule and the competitive binding molecule bind is not
in any way required in order to identify a competitive binding
molecule that binds to the same or substantially the same epitope
as the reference binding molecule.
[0659] In one aspect, the present disclosure provides a crystal of
RBD of the spike protein of SARS-CoV-2 in complex with an antibody.
In certain embodiments, the antibody complexed with the RBD in the
crystal is any antibody provided herein, or an antigen-binding
fragment thereof (e.g. an Fab fragment).
[0660] In some embodiment, the crystal provided herein comprises
Fab fragment of antibody P2B-2F6 in complex with RBD of the spike
protein of SARS-CoV-2. In some embodiment, the crystal consists of
a P2.sub.12.sub.12.sub.1 space group with unit cell dimensions of
a=70.23 .ANG., b=90.15 .ANG., and c=112.35 .ANG..
[0661] In some embodiment, the crystal provided herein comprises
Fab fragment of antibody P2C-1F11 in complex with RBD of the spike
protein of SARS-CoV-2. In some embodiment, the crystal has or
consists of a C121 space group with unit cell dimensions of
a=194.88 .ANG., b=85.39 .ANG., and c=58.51 .ANG..
[0662] In some embodiment, the crystal provided herein comprises
Fab fragment of antibody P22A-1D1 in complex with RBD of the spike
protein of SARS-CoV-2. In some embodiment, the crystal has or
consists of a C2 space group with unit cell dimensions of a=193.34
.ANG., b=86.60 .ANG., and c=57.16 .ANG..
[0663] In some embodiment, the crystal provided herein comprises
Fab fragment of antibody P5A-1D2 in complex with RBD of the spike
protein of SARS-CoV-2. In some embodiment, the crystal has or
consists of a C2 space group with unit cell dimensions of a=158.75
.ANG., b=67.51 .ANG., and c=154.37 .ANG..
[0664] In some embodiment, the crystal provided herein comprises
Fab fragment of antibody P5A-3C8 in complex with RBD of the spike
protein of SARS-CoV-2. In some embodiment, the crystal has or
consists of a P2.sub.12.sub.12.sub.1 space group with unit cell
dimensions of a=112.54 .ANG., b=171.57 .ANG., and c=54.87
.ANG..
[0665] X-ray crystallography analysis of the antibody bound to RBD
of the spike protein of SARS-CoV-2 can be used to determine
antibody epitopes. Epitopes may, in particular, be identified in
this way by determining residues on RBD of the spike protein of
SARS-CoV-2 within 4 .ANG. of an antibody paratope residue. In
another aspect, the present disclosure provides an isolated or
recombinant antibody or an antigen-binding fragment thereof, which
specifically binds to an epitope on RBD of spike protein of
SARS-CoV-2, wherein the epitope comprises at least one (at least
two, three, four, five, six, seven, eight, nine, ten, eleven, or
twelve) residues selected from K444, G446, G447, N448, Y449, N450,
L452, V483, E484, G485, F490 and S494, wherein the residue
numbering is according to SEQ ID NO: 134. In certain embodiments,
the epitope comprises Y449, L452, and F490. In certain embodiments,
the epitope comprises Y449, and G446. In certain embodiments, the
antibody or an antigen-binding fragment thereof provided herein has
a binding affinity (K.sub.d) to the RBD of spike protein of
SARS-CoV-2 of no more than 50 nM (e.g. no more than 40 nM, no more
than 30 nM, no more than 20 nM, no more than 10 nM, or no more than
5 nM), as measured by Surface Plasmon resonance (SPR).
[0666] In another aspect, the present disclosure provides an
isolated or recombinant antibody or an antigen-binding fragment
thereof, which specifically binds to an epitope on RBD of spike
protein of SARS-CoV-2, wherein the epitope comprises at least one
(at least two, three, four, five, six, seven, eight, nine, ten,
eleven, or twelve) residues selected from Y453, L455, F456, R457,
K458, 5459, N460, Y473, A475, G476, 5477, F486, N487, Y489, Q493,
G502, Y505, R403, T415, G416, K417, D420 and Y421, wherein the
residue numbering is according to SEQ ID NO: 134. In certain
embodiments, the epitope comprises at least one (at least two,
three, four, five, six, seven, eight, nine, ten, eleven, or twelve)
residues selected from Y453, L455, F456, R457, K458, S459, N460,
Y473, A475, G476, S477, F486, N487, Y489, Q493, G502 and Y505,
wherein the residue numbering is according to SEQ ID NO: 134. In
certain embodiments, the epitope comprises or further comprises at
least one (at least two, three, four, five, or six) residues
selected from R403, T415, G416, K417, D420 and Y421, wherein the
residue numbering is according to SEQ ID NO: 134. In certain
embodiments, the epitope comprises at least one (at least two,
three, four, five, six, seven, eight, nine, ten, eleven, twelve)
residues selected from T415, G416, K417, D420, Y421, L455, F456,
R457, K458, N460, Y473, A475, G476, S477, F486, N487, Y489 and
Q493, wherein the residue numbering is according to SEQ ID NO: 134.
In certain embodiments, the epitope comprises at least one (at
least two, three, four, five, six, seven, eight, nine, ten, eleven,
or twelve) residues selected from T415, G416, K417, D420, Y421,
Y453, L455, F456, R457, K458, N460, Y473, Q474, A475, G476, S477,
N487, Y489, Q493 and Y505, wherein the residue numbering is
according to SEQ ID NO: 134. In certain embodiments, the epitope
comprises at least one (at least two, three, four, five, six,
seven, eight, nine, ten, eleven, or twelve) residues selected from
T415, G416, K417, D420, Y421, Y453, L455, F456, R457, K458, N460,
Y473, A475, G476, S477, F486, N487, Y489 and Q493, wherein the
residue numbering is according to SEQ ID NO: 134. In certain
embodiments, the epitope comprises at least one (at least two,
three, four, five, six, seven, eight, nine, ten, eleven, or twelve)
residues selected from T415, G416, K417, D420, Y421, L455, F456,
R457, K458, N460, Y473, Q474, A475, G476, S477, F486, N487, Y489
and Q493, wherein the residue numbering is according to SEQ ID NO:
134. In certain embodiments, the epitope comprises at least one (at
least two, three, four, five, six or seven) residues selected from
L455, K458, Y473, A475, G476, S477 and N487. In certain
embodiments, the epitope comprises at least one (at least two,
three, four, five, six or seven) residues selected from T415, G416,
K417, D420, Y421, K458 and N460. In certain embodiments, the
epitope comprises at least one or at least two Y449, and G446. In
certain embodiments, the epitope comprises at least one (at least
two, three or four) residues selected from K417, Y421, L455 and
F456. In certain embodiments, the epitope comprises at least one
(at least two, three, or four) residues selected from F456, N487,
Y489 and Q493. In certain embodiments, the epitope comprises L455.
In certain embodiments, the epitope comprises at least one or at
least two residues selected from Y421 and D420. In certain
embodiments, the epitope comprises Y421. In certain embodiments,
the epitope comprises Y505. In certain embodiments, the epitope
comprises Y421A and F456A, wherein the residue numbering is
according to SEQ ID NO: 134. In certain embodiments, the epitope
comprises T415A, Y473A, and N487A, wherein the residue numbering is
according to SEQ ID NO: 134. In certain embodiments, the epitope
comprises K417A, D420A, L455A, R457A, N460A, and Y489A, wherein the
residue numbering is according to SEQ ID NO: 134. In certain
embodiments, the epitope comprises T415A, Y421A, L455A, F456A,
R457A, Y473A, N487A, Y489A, and Y505A, wherein the residue
numbering is according to SEQ ID NO: 134. In certain embodiments,
the antibody or an antigen-binding fragment thereof provided herein
has a binding affinity (K.sub.d) to the RBD of spike protein of
SARS-CoV-2 of no more than 50 nM (e.g. no more than 40 nM, no more
than 30 nM, no more than 20 nM, or no more than 10 nM, or no more
than 5 nM), as measured by Surface Plasmon resonance (SPR).
[0667] In an aspect, the present disclosure provides a
computer-implemented method for causing a display of a graphical
three-dimensional representation of the structure of a portion of a
crystal of RBD of the spike protein of SARS-CoV-2 in complex with
an anti-SARS-CoV-2 antibody or an antigen-binding fragment thereof
provided herein, wherein the method comprises: causing said display
of said graphical three-dimensional representation by a computer
system programmed with instructions for transforming structure
coordinates into said graphical three-dimensional representation of
said structure and for displaying said graphical three-dimensional
representation, wherein said graphical three-dimensional
representation is generated by transforming said structure
coordinates into said graphical three-dimensional representation of
said structure, wherein said structure coordinates comprise
structure coordinates of the backbone atoms of the portion of the
crystal, wherein the portion of the crystal comprises a RBD binding
site, and wherein the crystal has the space group symmetry
P2.sub.12.sub.12.sub.1 or C121.
[0668] In another aspect, the present disclosure provides a
machine-readable data storage medium comprising a data storage
material encoded with machine-readable instructions for: (a)
transforming data into a graphical three-dimensional representation
for the structure of a portion of a crystal of RBD of the spike
protein of SARS-CoV-2 in complex with an anti-SARS-CoV-2 antibody
or an antigen-binding fragment thereof provided herein; and (b)
causing the display of said graphical three-dimensional
representation; wherein said data comprise structure coordinates of
the backbone atoms of the amino acids defining a RBD binding site;
and wherein the crystal or structural homolog has the space group
symmetry P2.sub.12.sub.12.sub.1 or C121.
[0669] In another aspect, the present disclosure provides a
computer system for displaying a three-dimensional graphical
representation for the structure of a portion of a crystal of RBD
of the spike protein of SARS-CoV-2 in complex with an
anti-SARS-CoV-2 antibody or an antigen-binding fragment thereof as
provided herein, comprising: (a) a machine-readable data storage
medium comprising a data storage material encoded with
machine-readable data, wherein said data comprise structure
coordinates of the backbone atoms of the amino acids defining a RBD
binding site, wherein the crystal has the space group symmetry
P2.sub.12.sub.12.sub.1 or C121; (b) a working memory; (c) a central
processing unit coupled to said working memory and to said
machine-readable data storage medium for processing said
machine-readable data into sad three-dimensional graphical
representation; and (d) a display coupled to said central
processing unit for displaying said three-dimensional graphical
representation.
[0670] For the above listed aspects, in certain embodiments, the
RBD comprises an amino acid sequence as shown in SEQ ID NO: 124. In
certain embodiments, the antibody comprises a pair of heavy chain
variable region and light chain variable region as listed in Table
2, or the homologous sequence thereof (e.g. having at least 80%
sequence identity). In certain embodiments, the antibody comprises:
a) a heavy chain variable region of SEQ ID NO: 47 and a light chain
variable region of SEQ ID NO: 48; orb) a heavy chain variable
region of SEQ ID NO: 111 and a light chain variable region of SEQ
ID NO: 112; or c) a heavy chain variable region of SEQ ID NO: 432
and a light chain variable region of SEQ ID NO: 433; or d) a heavy
chain variable region of SEQ ID NO: 242 and a light chain variable
region of SEQ ID NO: 243; or e) a heavy chain variable region of
SEQ ID NO: 232 and a light chain variable region of SEQ ID NO: 233.
In certain embodiments, the structure coordinates comprise the
structure coordinates of the backbone atoms of the amino acid
residues corresponding to K444, G446, G447, N448, Y449, N450, L452,
V483, E484, G485, F490 and/or S494 of the RBD, wherein the residue
numbering is according to SEQ ID NO: 134. In certain embodiments,
the structure coordinates comprise the structure coordinates of the
backbone atoms of the amino acid residues corresponding to Y453,
L455, F456, R457, K458, S459, N460, Y473, A475, G476, S477, F486,
N487, Y489, Q493, G502, Y505, R403, T415, G416, K417, D420 and/or
Y421 of the RBD, wherein the residue numbering is according to SEQ
ID NO: 134. In certain embodiments, the structure coordinates
comprise the structure coordinates of the backbone atoms of the
amino acid residues corresponding to T415, G416, K417, D420, Y421,
L455, F456, R457, K458, N460, Y473, A475, G476, S477, F486, N487,
Y489 and/or Q493 of the RBD, wherein the residue numbering is
according to SEQ ID NO: 134. In certain embodiments, the structure
coordinates comprise the structure coordinates of the backbone
atoms of the amino acid residues corresponding to T415, G416, K417,
D420, Y421, Y453, L455, F456, R457, K458, N460, Y473, Q474, A475,
G476, S477, N487, Y489, Q493 and/or Y505 of the RBD, wherein the
residue numbering is according to SEQ ID NO: 134. In certain
embodiments, the structure coordinates comprise the structure
coordinates of the backbone atoms of the amino acid residues
corresponding to T415, G416, K417, D420, Y421, Y453, L455, F456,
R457, K458, N460, Y473, A475, G476, 5477, F486, N487, Y489 and/or
Q493 of the RBD, wherein the residue numbering is according to SEQ
ID NO: 134. In certain embodiments, the structure coordinates
comprise the structure coordinates of the backbone atoms of the
amino acid residues corresponding to T415, G416, K417, D420, Y421,
L455, F456, R457, K458, N460, Y473, Q474, A475, G476, S477, F486,
N487, Y489 and/or Q493 of the RBD, wherein the residue numbering is
according to SEQ ID NO: 134.
[0671] In another aspect, the present disclosure provides a method
of screening for molecules that may be a binding molecule of RBD of
the spike protein of SARS-CoV-2, comprising: (a) computationally
screening agents against a three-dimensional model to identify
potential binding molecules of the RBD; wherein the
three-dimensional model comprises a three-dimensional model of at
least a portion of a crystal of RBD of the spike protein of
SARS-CoV-2 in complex with an anti-SARS-CoV-2 antibody or an
antigen-binding fragment thereof; wherein the three dimensional
model is generated from at least a portion of the structure
coordinates of the crystal by a computer algorithm for generating a
three-dimensional model of the crystal useful for identifying
agents that are potential binding molecules of the RBD.
[0672] In certain embodiments, the crystal comprises a polypeptide
comprising an amino acid sequence SEQ ID NO: 124, or a homologous
sequence thereof, for example derived from a mutant SARS-CoV-2. In
certain embodiments, the crystal further comprises an antibody or
antigen-binding fragment thereof comprising a pair of heavy chain
variable region and light chain variable region as listed in Table
2, or the homologous sequence thereof (e.g. having at least 80%
sequence identity). In certain embodiments, the crystal further
comprises an antibody or antigen-binding fragment thereof
comprising: a) a heavy chain variable region of SEQ ID NO: 47 and a
light chain variable region of SEQ ID NO: 48, or b) a heavy chain
variable region of SEQ ID NO: 111 and a light chain variable region
of SEQ ID NO: 112, wherein the crystal diffracts x-rays for the
determination of atomic coordinates to a resolution of 5 .ANG. or
better.
[0673] A method for obtaining structural information about a
molecule or molecular complex comprising applying at least a
portion of the structure coordinates of a RBD of the spike protein
of SARS-CoV-2 in complex with an anti-SARS-CoV-2 antibody or an
antigen-binding fragment thereof provided herein, to an X-ray
diffraction pattern of the molecule or molecular complex's crystal
structure to cause the generation of a three-dimensional electron
density map of at least a portion of the molecule or molecular
complex. In certain embodiments, the crystal comprises a
polypeptide comprising an amino acid sequence SEQ ID NO: 124, or a
homologous sequence thereof, for example derived from a mutant
SARS-CoV-2. In certain embodiments, the crystal further comprises
an antibody or antigen-binding fragment thereof comprising a pair
of heavy chain variable region and light chain variable region as
listed in Table 2, or the homologous sequence thereof (e.g. having
at least 80% sequence identity). In certain embodiments, the
crystal further comprises an antibody or antigen-binding fragment
thereof comprising: a) a heavy chain variable region of SEQ ID NO:
47 and a light chain variable region of SEQ ID NO: 48, orb) a heavy
chain variable region of SEQ ID NO: 111 and a light chain variable
region of SEQ ID NO: 112, wherein the crystal diffracts x-rays for
the determination of atomic coordinates to a resolution of 5 .ANG.
or better.
[0674] Conjugates
[0675] In some embodiments, the anti-SARS-CoV-2 antibodies or
antigen-binding fragments thereof further comprise one or more
conjugate moieties. A conjugate moiety is a moiety that can be
attached to the antibodies or antigen-binding fragments thereof
either directly or via a linker or through another conjugate
moiety. It is contemplated that a variety of conjugate moieties may
be linked to the antibodies or antigen-binding fragments thereof
provided herein (see, for example, "Conjugate Vaccines",
Contributions to Microbiology and Immunology, J. M. Cruse and R. E.
Lewis, Jr. (eds.), Carger Press, New York, (1989)). These conjugate
moieties may be linked to the antibodies or antigen-binding
fragments thereof by covalent binding, affinity binding,
intercalation, coordinate binding, complexation, association,
blending, or addition, among other methods.
[0676] In certain embodiments, the antibodies or antigen-binding
fragments thereof provided herein may be engineered to contain
specific sites outside the epitope binding portion that may be
utilized for binding to one or more conjugate moieties. For
example, such a site may include one or more reactive amino acid
residues, such as for example cysteine or histidine residues, to
facilitate covalent linkage to a conjugate moiety.
[0677] Examples of such conjugate moieties include but are not
limited to, therapeutic agent, a radioactive isotope, a detectable
label, a pharmacokinetic modifying moiety, or a purifying moiety.
In some embodiments, the conjugate moiety comprises a
clearance-modifying agent (e.g. a polymer such as PEG which extends
half-life), a chemotherapeutic agent, a toxin, a radioactive
isotope, a lanthanide, a detectable label (e.g. a luminescent
label, a fluorescent label, an enzyme-substrate label), a
DNA-alkylator, a topoisomerase inhibitor, a tubulin-binder, a
purification moiety or other anticancer drugs.
[0678] Examples of detectable label may include a fluorescent
labels (e.g. fluorescein, rhodamine, dansyl, phycoerythrin, or
Texas Red), enzyme-substrate labels (e.g. horseradish peroxidase,
alkaline phosphatase, luceriferases, glucoamylase, lysozyme,
saccharide oxidases or .beta.-D-galactosidase), radioisotopes (e.g.
.sup.123I, .sup.124I, .sup.125I, .sup.131I, .sup.35S, .sup.3H,
.sup.111In, .sup.112In, .sup.14C, .sup.64Cu, .sup.67Cu, .sup.86Y,
.sup.88Y, .sup.90Y, .sup.177Lu, .sup.211At, .sup.186Re, .sup.188Re,
.sup.153Sm, .sup.212Bi, and .sup.32P, other lanthanides),
luminescent labels, chromophoric moieties, digoxigenin,
biotin/avidin, DNA molecules or gold for detection.
[0679] In certain embodiments, the conjugate moiety can be a
clearance-modifying agent which helps increase half-life of the
antibody. Illustrative example include water-soluble polymers, such
as PEG, carboxymethylcellulose, dextran, polyvinyl alcohol,
polyvinyl pyrrolidone, copolymers of ethylene glycol/propylene
glycol, and the like. The polymer may be of any molecular weight,
and may be branched or unbranched. The number of polymers attached
to the antibody may vary, and if more than one polymer are
attached, they can be the same or different molecules. In certain
embodiments, the conjugate moiety can be a purification moiety such
as a magnetic bead. In certain embodiments, the antibodies or
antigen-binding fragments thereof provided herein is used as a base
for a conjugate.
[0680] Polynucleotides and Recombinant Methods
[0681] The present disclosure provides isolated polynucleotides
that encode the anti-SARS-CoV-2 antibodies or antigen-binding
fragments thereof provided herein. DNA encoding the monoclonal
antibody is readily isolated, e.g., from B cells, and sequenced
using conventional procedures (e.g. by using oligonucleotide probes
that are capable of binding specifically to genes encoding the
heavy and light chains of the antibody). The encoding DNA may also
be obtained by synthetic methods.
[0682] The isolated polynucleotide that encodes the anti-SARS-CoV-2
antibodies or antigen-binding fragments thereof can be inserted
into a vector for further cloning (amplification of the DNA) or for
expression (i.e., expression vector), using recombinant techniques
known in the art. Many vectors are available. The vector components
generally include, but are not limited to, one or more of the
following: a signal sequence, an origin of replication, one or more
marker genes, an enhancer element, a promoter (e.g. SV40, CMV,
EF-1.alpha.), and a transcription termination sequence.
[0683] The present disclosure provides vectors comprising the
isolated polynucleotide provided herein. In certain embodiments,
the polynucleotide provided herein encodes the antibodies or
antigen-binding fragments thereof, at least one promoter (e.g.
SV40, CMV, EF-1.alpha.) operably linked to the nucleic acid
sequence, and at least one selection marker. Examples of vectors
include, but are not limited to, retrovirus (including lentivirus),
adenovirus, adeno-associated virus, herpesvirus (e.g. herpes
simplex virus), poxvirus, baculovirus, papillomavirus, papovavirus
(e.g. SV40), lambda phage, and M13 phage, plasmid pcDNA3.3,
pMD18-T, pOptivec, pCMV, pEGFP, pIRES, pQD-Hyg-GSeu, pALTER, pBAD,
pcDNA, pCal, pL, pET, pGEMEX, pGEX, pCI, pEGFT, pSV2, pFUSE,
pVITRO, pVIVO, pMAL, pMONO, pSELECT, pUNO, pDUO, Psg5L, pBABE,
pWPXL, pBI, p15TV-L, pPro18, pTD, pRS10, pLexA, pACT2.2,
pCMV-SCRIPT.RTM., pCDM8, pCDNA1.1/amp, pcDNA3.1, pRc/RSV, PCR 2.1,
pEF-1, pFB, pSG5, pXT1, pCDEF3, pSVSPORT, pEF-Bos etc.
[0684] Vectors comprising the polynucleotide sequence encoding the
antibody or antigen-binding fragment thereof can be introduced to a
host cell for cloning or gene expression. Suitable host cells for
cloning or expressing the DNA in the vectors herein are the
prokaryote, yeast, or higher eukaryote cells described above.
Suitable prokaryotes for this purpose include eubacteria, such as
Gram-negative or Gram-positive organisms, for example,
Enterobacteriaceae such as Escherichia, e.g. E. coli, Enterobacter,
Envinia, Klebsiella, Proteus, Salmonella, e.g. Salmonella
typhimurium, Serratia, e.g. Serratia marcescans, and Shigella, as
well as Bacilli such as B. subtilis and B. licheniformis,
Pseudomonas such as P. aeruginosa, and Streptomyces.
[0685] In addition to prokaryotes, eukaryotic microbes such as
filamentous fungi or yeast are suitable cloning or expression hosts
for anti-SARS-CoV-2 antibody-encoding vectors. Saccharomyces
cerevisiae, or common baker's yeast, is the most commonly used
among lower eukaryotic host microorganisms. However, a number of
other genera, species, and strains are commonly available and
useful herein, such as Schizosaccharomyces pombe; Kluyveromyces
hosts such as, e.g. K. lactis, K. fragilis (ATCC 12,424), K.
bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K. waltii
(ATCC 56,500), K. drosophilarum (ATCC 36,906), K. thermotolerans,
and K. marxianus; yarrowia (EP 402,226); Pichia pastoris (EP
183,070); Candida; Trichoderma reesia (EP 244,234); Neurospora
crassa; Schwanniomyces such as Schwanniomyces occidentalis; and
filamentous fungi such as, e.g. Neurospora, Penicillium,
Tolypocladium, and Aspergillus hosts such as A. nidulans and A.
niger.
[0686] Suitable host cells for the expression of glycosylated
antibodies or antigen-fragment thereof provided herein are derived
from multicellular organisms. Examples of invertebrate cells
include plant and insect cells. Numerous baculoviral strains and
variants and corresponding permissive insect host cells from hosts
such as Spodoptera frugiperda (caterpillar), Aedes aegypti
(mosquito), Aedes albopictus (mosquito), Drosophila melanogaster
(fruiffly), and Bombyx mori have been identified. A variety of
viral strains for transfection are publicly available, e.g. the L-1
variant of Autographa californica NPV and the Bm-5 strain of Bombyx
mori NPV, and such viruses may be used as the virus herein
according to the present invention, particularly for transfection
of Spodoptera frupperda cells. Plant cell cultures of cotton, corn,
potato, soybean, petunia, tomato, and tobacco can also be utilized
as hosts.
[0687] However, interest has been greatest in vertebrate cells, and
propagation of vertebrate cells in culture (tissue culture) has
become a routine procedure. Examples of useful mammalian host cell
lines are monkey kidney CV1 line transformed by SV40 (COS-7, ATCC
CRL 1651); human embryonic kidney line (293 or 293 cells subcloned
for growth in suspension culture, Graham et al., J. Gen Virol.
36:59 (1977)); baby hamster kidney cells (BHK, ATCC CCL 10);
Chinese hamster ovary cells/-DHFR (CHO, Urlaub et al., Proc. Natl.
Acad. Sci. USA 77:4216 (1980)); mouse sertoli cells (TM4, Mather,
Biol. Reprod. 23:243-251 (1980)); monkey kidney cells (CV1 ATCC CCL
70); African green monkey kidney cells (VERO-76, ATCC CRL-1587);
human cervical carcinoma cells (HELA, ATCC CCL 2); canine kidney
cells (MDCK, ATCC CCL 34); buffalo rat liver cells (BRL 3A, ATCC
CRL 1442); human lung cells (W138, ATCC CCL 75); human liver cells
(Hep G2, HB 8065); mouse mammary tumor (MMT 060562, ATCC CCL51);
TRI cells (Mather et al., Annals N.Y. Acad. Sci. 383:44-68 (1982));
MRC 5 cells; FS4 cells; and a human hepatoma line (Hep G2). In some
embodiments, the host cell is a mammalian cultured cell line, such
as CHO, BHK, NS0, 293 and their derivatives.
[0688] Host cells are transformed with the above-described
expression or cloning vectors for anti-SARS-CoV-2 antibody
production and cultured in conventional nutrient media modified as
appropriate for inducing promoters, selecting transformants, or
amplifying the genes encoding the desired sequences. In another
embodiment, the antibody may be produced by homologous
recombination known in the art. In certain embodiments, the host
cell is capable of producing the antibody or antigen-binding
fragment thereof provided herein.
[0689] The present disclosure also provides a method of expressing
the antibody or an antigen-binding fragment thereof provided
herein, comprising culturing the host cell provided herein under
the condition at which the vector of the present disclosure is
expressed. The host cells used to produce the antibodies or
antigen-binding fragments thereof provided herein may be cultured
in a variety of media. Commercially available media such as Ham's
F10 (Sigma), Minimal Essential Medium (MEM), (Sigma), RPMI-1640
(Sigma), and Dulbecco's Modified Eagle's Medium (DMEM), Sigma) are
suitable for culturing the host cells. In addition, any of the
media described in Ham et al., Meth. Enz. 58:44 (1979), Barnes et
al., Anal. Biochem. 102:255 (1980), U.S. Pat. Nos. 4,767,704;
4,657,866; 4,927,762; 4,560,655; or 5,122,469; WO 90/03430; WO
87/00195; or U.S. Pat. Re. 30,985 may be used as culture media for
the host cells. Any of these media may be supplemented as necessary
with hormones and/or other growth factors (such as insulin,
transferrin, or epidermal growth factor), salts (such as sodium
chloride, calcium, magnesium, and phosphate), buffers (such as
HEPES), nucleotides (such as adenosine and thymidine), antibiotics
(such as GENTAMYCIN.TM. drug), trace elements (defined as inorganic
compounds usually present at final concentrations in the micromolar
range), and glucose or an equivalent energy source. Any other
necessary supplements may also be included at appropriate
concentrations that would be known to a person skilled in the art.
The culture conditions, such as temperature, pH, and the like, are
those previously used with the host cell selected for expression,
and will be apparent to a person skilled in the art.
[0690] When using recombinant techniques, the antibody can be
produced intracellularly, in the periplasmic space, or directly
secreted into the medium. If the antibody is produced
intracellularly, as a first step, the particulate debris, either
host cells or lysed fragments, is removed, for example, by
centrifugation or ultrafiltration. Carter et al., Bio/Technology
10:163-167 (1992) describe a procedure for isolating antibodies
which are secreted to the periplasmic space of E. coli. Briefly,
cell paste is thawed in the presence of sodium acetate (pH 3.5),
EDTA, and phenylmethylsulfonylfluoride (PMSF) over about 30 min.
Cell debris can be removed by centrifugation. Where the antibody is
secreted into the medium, supernatants from such expression systems
are generally first concentrated using a commercially available
protein concentration filter, for example, an Amicon or Millipore
Pellicon ultrafiltration unit. A protease inhibitor such as PMSF
may be included in any of the foregoing steps to inhibit
proteolysis and antibiotics may be included to prevent the growth
of adventitious contaminants.
[0691] The anti-SARS-CoV-2 antibodies or antigen-binding fragments
thereof prepared from the cells can be purified using, for example,
hydroxylapatite chromatography, gel electrophoresis, dialysis,
DEAE-cellulose ion exchange chromatography, ammonium sulfate
precipitation, salting out, and affinity chromatography, with
affinity chromatography being the preferred purification
technique.
[0692] In certain embodiments, Protein A immobilized on a solid
phase is used for immunoaffinity purification of the antibody and
antigen-binding fragment thereof. The suitability of protein A as
an affinity ligand depends on the species and isotype of any
immunoglobulin Fc domain that is present in the antibody. Protein A
can be used to purify antibodies that are based on human gammal,
gamma2, or gamma4 heavy chains (Lindmark et al., J. Immunol. Meth.
62:1-13 (1983)). Protein G is recommended for all mouse isotypes
and for human gamma3 (Guss et al., EMBO J. 5:1567 1575 (1986)). The
matrix to which the affinity ligand is attached is most often
agarose, but other matrices are available. Mechanically stable
matrices such as controlled pore glass or
poly(styrenedivinyl)benzene allow for faster flow rates and shorter
processing times than can be achieved with agarose. Where the
antibody comprises a CH3 domain, the Bakerbond ABX.TM. resin (J. T.
Baker, Phillipsburg, N.J.) is useful for purification. Other
techniques for protein purification such as fractionation on an
ion-exchange column, ethanol precipitation, Reverse Phase HPLC,
chromatography on silica, chromatography on heparin crosslinked,
beaded-form of agarose SEPHAROSE.TM. (trademark of GE Healthcare)
chromatography on an anion or cation exchange resin (such as a
polyaspartic acid column), chromatofocusing, SDS-PAGE, and ammonium
sulfate precipitation are also available depending on the antibody
to be recovered.
[0693] Following any preliminary purification step(s), the mixture
comprising the antibody of interest and contaminants may be
subjected to low pH hydrophobic interaction chromatography using an
elution buffer at a pH between about 2.5-4.5, preferably performed
at low salt concentrations (e.g. from about 0-0.25M salt).
[0694] Pharmaceutical Composition
[0695] The present disclosure further provides pharmaceutical
compositions comprising the anti-SARS-CoV-2 antibodies or
antigen-binding fragments thereof and one or more pharmaceutically
acceptable carriers.
[0696] The present disclosure further provides a pharmaceutical
composition comprising at least one or more of the modified
antibody or an antigen-binding fragment thereof disclosed herein,
at least one nucleic acid encoding the modified antibody or the
antigen-binding fragment thereof, or a combination thereof, and one
or more pharmaceutically acceptable carriers.
[0697] In some embodiments, the pharmaceutical composition
comprises a combination of two or more antibodies or the antigen
binding fragments of the present disclosure. In some embodiments,
the pharmaceutical composition comprises a combination of two or
more monoclonal antibodies, each of which comprises heavy chain CDR
sequences and light chain CDR sequences derived from an antibody
selected from the group consisting of P2A-1A8, P2A-1A9, P2B-2G11,
P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4, P2C-1A3, P2C-1C8, P2C- 1C10,
P2C-1D5, P2C-1F11, P2B-1G5, P2B-1A1, P2C-1D7, P2B-1A10, P2B-1D9,
P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2, P5A-2F11,
P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6, P5A-1B8,
P5A-1B9, P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9, P5A-2H3, P5A-3A1,
P5A-3A6, P5A-3B4, P5A-3C12, and P22A-1D1. In some embodiments, the
pharmaceutical composition comprises a first antibody comprising
heavy chain CDR sequences and light chain CDR sequences derived
from P2C-1F11, and a second antibody comprising heavy chain CDR
sequences and light chain CDR sequences derived from antibody
P2B-2F6.
[0698] In some embodiments, the two or more antibodies or the
antigen binding fragments thereof bind to different epitopes in RBD
of spike protein of SARS-CoV-2. In certain embodiments, the
pharmaceutical composition comprises a first antibody which
comprises P2C-1F11 or an antigen binding fragment thereof, and a
second antibody which is selected from the group consisting of
P2C-1A3, P2C-1C10, P2B-2F6, P2B-1G5, and P2A-1B3, or an antigen
binding fragment thereof. In certain embodiments, the
pharmaceutical composition comprises a first antibody which
comprises P2C-1A3 or an antigen binding fragment thereof, and a
second antibody which is selected from the group consisting of
P5A-3C8, P5A-1D2, P22A-1D1, P2C-1F11, and P2A-1B3, or an antigen
binding fragment thereof In certain embodiments, the pharmaceutical
composition comprises a first antibody which comprises P2B-2F6 or
an antigen binding fragment thereof, and a second antibody selected
from the group consisting of P2C-1C10, P2C-1F11, P2B-1G5, and
P2A-1B3, or an antigen binding fragment thereof. In certain
embodiments, the pharmaceutical composition comprises a first
antibody which comprises P2A-1B3 or an antigen binding fragment
thereof, and a second antibody selected from the group consisting
of P5A-3C8, P5A-1D2, P22A-1D1, P2C-1A3, P2C-1C10, P2C-1F11,
P2B-2F6, and P2A-1A10, or an antigen binding fragment thereof. In
some embodiments, the pharmaceutical composition comprises a first
antibody which comprises P2C-1C10 or an antigen binding fragment
thereof, and a second antibody selected from the group consisting
of P5A-3C8, P5A-1D2, P22A-1D1, P2C-1A3, P2C-1F11, and P2A-1B3, or
an antigen binding fragment thereof.
[0699] The present disclosure further provides pharmaceutical
compositions comprising the polynucleotides encoding the
anti-SARS-CoV-2 antibodies or the antigen-binding fragments
thereof, and one or more pharmaceutically acceptable carriers. The
present disclosure further provides pharmaceutical compositions
comprising the polynucleotides encoding the combination of the two
or more anti-SARS-CoV-2 antibodies or the antigen-binding fragments
thereof, and one or more pharmaceutically acceptable carriers.
[0700] The present disclosure further provides pharmaceutical
compositions comprising an expression vector comprising the
polynucleotides encoding the one or more of anti-SARS-CoV-2
antibodies or the antigen-binding fragments thereof, and one or
more pharmaceutically acceptable carriers.
[0701] In certain embodiments, the expression vector comprises a
viral vector or a non-viral vector. Examples of viral vectors
include, without limitation, adeno-associated virus (AAV) vector,
lentivirus vector, retrovirus vector, and adenovirus vector.
Examples of non-viral vectors include, without limitation, naked
DNA, plasmid, exosome, mRNA, and so on. In certain embodiments, the
expression vector is suitable for gene therapy in human. Suitable
vectors for gene therapy include, for example, adeno-associated
virus (AAV), or adenovirus vector. In certain embodiments, the
expression vector comprises a DNA vector or a RNA vector. In
certain embodiments, the pharmaceutically acceptable carriers are
polymeric excipients, such as without limitation, microspheres,
microcapsules, polymeric micelles and dendrimers. The
polynucleotides, or polynucleotide vectors of the present
disclosure may be encapsulated, adhered to, or coated on the
polymer-based components by methods known in the art (see for
example, W. Heiser, Nonviral gene transfer techniques, published by
Humana Press, 2004; U.S. Pat. No. 6,025,337; Advanced Drug Delivery
Reviews, 57(15): 2177-2202 (2005)).
[0702] In some embodiments, the pharmaceutical composition further
comprises a second bioactive agent, such as a second therapeutic
agent or a second prophylactic agent.
[0703] Pharmaceutical acceptable carriers for use in the
pharmaceutical compositions disclosed herein may include, for
example, pharmaceutically acceptable liquid, gel, or solid
carriers, aqueous vehicles, nonaqueous vehicles, antimicrobial
agents, isotonic agents, buffers, antioxidants, anesthetics,
suspending/dispending agents, sequestering or chelating agents,
diluents, adjuvants, excipients, or non-toxic auxiliary substances,
other components known in the art, or various combinations
thereof.
[0704] Suitable components may include, for example, antioxidants,
fillers, binders, disintegrants, buffers, preservatives,
lubricants, flavorings, thickeners, coloring agents, emulsifiers or
stabilizers such as sugars and cyclodextrins. Suitable antioxidants
may include, for example, methionine, ascorbic acid, EDTA, sodium
thiosulfate, platinum, catalase, citric acid, cysteine,
thioglycerol, thioglycolic acid, thiosorbitol, butylated
hydroxanisol, butylated hydroxytoluene, and/or propyl gallate. As
disclosed herein, inclusion of one or more antioxidants such as
methionine in a composition comprising an antibody or
antigen-binding fragment thereof and conjugates provided herein
decreases oxidation of the antibody or antigen-binding fragment
thereof. This reduction in oxidation prevents or reduces loss of
binding affinity, thereby improving antibody stability and
maximizing shelf-life. Therefore, in certain embodiments,
pharmaceutical compositions are provided that comprise one or more
antibodies or antigen-binding fragments thereof as disclosed herein
and one or more antioxidants such as methionine. Further provided
are methods for preventing oxidation of, extending the shelf-life
of, and/or improving the efficacy of an antibody or antigen-binding
fragment provided herein by mixing the antibody or antigen-binding
fragment with one or more antioxidants such as methionine.
[0705] To further illustrate, pharmaceutical acceptable carriers
may include, for example, aqueous vehicles such as sodium chloride
injection, Ringer's injection, isotonic dextrose injection, sterile
water injection, or dextrose and lactated Ringer's injection,
nonaqueous vehicles such as fixed oils of vegetable origin,
cottonseed oil, corn oil, sesame oil, or peanut oil, antimicrobial
agents at bacteriostatic or fungistatic concentrations, isotonic
agents such as sodium chloride or dextrose, buffers such as
phosphate or citrate buffers, antioxidants such as sodium
bisulfate, local anesthetics such as procaine hydrochloride,
suspending and dispersing agents such as sodium
carboxymethylcelluose, hydroxypropyl methylcellulose, or
polyvinylpyrrolidone, emulsifying agents such as nonionic
surfactant Polysorbate 80 (TWEEN.RTM.-80, TWEEN is a registered
trademark of CRODA AMERICAS LLC), sequestering or chelating agents
such as EDTA (ethylenediaminetetraacetic acid) or EGTA (ethylene
glycol tetraacetic acid), ethyl alcohol, polyethylene glycol,
propylene glycol, sodium hydroxide, hydrochloric acid, citric acid,
or lactic acid. Antimicrobial agents utilized as carriers may be
added to pharmaceutical compositions in multiple-dose containers
that include phenols or cresols, mercurials, benzyl alcohol,
chlorobutanol, methyl and propyl p-hydroxybenzoic acid esters,
thimerosal, benzalkonium chloride and benzethonium chloride.
Suitable excipients may include, for example, water, saline,
dextrose, glycerol, or ethanol. Suitable non-toxic auxiliary
substances may include, for example, wetting or emulsifying agents,
pH buffering agents, stabilizers, solubility enhancers, or agents
such as sodium acetate, sorbitan monolaurate, triethanolamine
oleate, or cyclodextrin.
[0706] The pharmaceutical compositions can be a liquid solution,
suspension, emulsion, pill, capsule, tablet, sustained release
formulation, or powder. Oral formulations can include standard
carriers such as pharmaceutical grades of mannitol, lactose,
starch, magnesium stearate, polyvinyl pyrollidone, sodium
saccharine, cellulose, magnesium carbonate, etc.
[0707] The form of pharmaceutical compositions depends on a number
of criteria, including, but not limited to, route of
administration, extent of disease, or dose to be administered. The
pharmaceutical compositions can be formulated for intravenous,
oral, nasal, rectal, percutaneous, or intramuscular administration.
For example, dosage forms for intravenous administration, may be
formulated as lyophilized powder or fluid formulation; dosage forms
for nasal administration may conveniently be formulated as
aerosols, solutions, drops, gels or dry powders. In accordance to
the desired route of administration, the pharmaceutical
compositions can be formulated in the form of tablets, capsule,
pill, dragee, powder, granule, sachets, cachets, lozenges,
suspensions, emulsions, solutions, syrups, aerosols (as a solid or
in a liquid medium), spray, inhalant, or suppository.
[0708] In certain embodiments, the pharmaceutical compositions are
formulated into an injectable composition. The injectable
pharmaceutical compositions may be prepared in any conventional
form, such as for example liquid solution, suspension, emulsion, or
solid forms suitable for generating liquid solution, suspension, or
emulsion. Preparations for injection may include sterile and/or
non-pyretic solutions ready for injection, sterile dry soluble
products, such as lyophilized powders, ready to be combined with a
solvent just prior to use, including hypodermic tablets, sterile
suspensions ready for injection, sterile dry insoluble products
ready to be combined with a vehicle just prior to use, and sterile
and/or non-pyretic emulsions. The solutions may be either aqueous
or nonaqueous.
[0709] In certain embodiments, unit-dose parenteral preparations
are packaged in an ampoule, a vial or a syringe with a needle. All
preparations for parenteral administration should be sterile and
not pyretic, as is known and practiced in the art.
[0710] In certain embodiments, a sterile, lyophilized powder is
prepared by dissolving an antibody or antigen-binding fragment as
disclosed herein in a suitable solvent. The solvent may contain an
excipient which improves the stability or other pharmacological
components of the powder or reconstituted solution, prepared from
the powder. Excipients that may be used include, but are not
limited to, water, dextrose, sorbital, fructose, corn syrup,
xylitol, glycerin, glucose, sucrose or other suitable agent. The
solvent may contain a buffer, such as citrate, sodium or potassium
phosphate or other such buffer known to a person skilled in the art
at, in one embodiment, about neutral pH. Subsequent sterile
filtration of the solution followed by lyophilization under
standard conditions known to a person skilled in the art provides a
desirable formulation. In one embodiment, the resulting solution
will be apportioned into vials for lyophilization. Each vial can
contain a single dosage or multiple dosages of the anti-SARS-CoV-2
antibody or antigen-binding fragment thereof or composition
thereof. Overfilling vials with a small amount above that needed
for a dose or set of doses (e.g. about 10%) is acceptable so as to
facilitate accurate sample withdrawal and accurate dosing. The
lyophilized powder can be stored under appropriate conditions, such
as at about 4.degree. C. to room temperature.
[0711] Reconstitution of a lyophilized powder with water for
injection provides a formulation for use in parenteral
administration. In one embodiment, for reconstitution the sterile
and/or non-pyretic water or other liquid suitable carrier is added
to lyophilized powder. The precise amount depends upon the selected
therapy being given, and can be empirically determined.
[0712] The pharmaceutical composition disclosed herein can comprise
the modified antibody or an antigen-binding fragment thereof at a
concentration in a range of from a concentration in a range of from
10 mg/mL to 150 mg/mL. The concentration of the modified antibody
or an antigen-binding fragment thereof can be determined based on
total protein concentration, antibody specific protein
concentration, or a combination thereof. Typical measurement method
for measuring protein concentrations known to those skilled in the
art can be suitable.
[0713] In some embodiments, the pharmaceutical composition can be
configured to be administered to a subject via intravenous
injection (IV), intramuscular injection (IM), subcutaneous (SC)
injection, or a combination thereof.
[0714] In some embodiments, the pharmaceutical composition can be
configured for preventing a disease in a person having no symptoms
or free from known infections of the SARS-CoV-2, or treatment of a
patient being a symptomatic non-hospitalized person of any age or
an adult with COVID-19 caused by SARS-CoV-2 infection, aged 60
years and older, any age having at least one of the following
conditions selected from smoking, exogenous or endogenous
immunosuppression having HIV infection with CD4 count <200
cells/mm.sup.3, receiving corticosteroids equivalent to prednisone
.gtoreq.20 mg daily for at least 14 consecutive days within 30 days
prior to be administered with the pharmaceutical composition,
receiving one or more biologics therapeutical agents, one or more
immunomodulators, cancer chemotherapy within 90 days prior to be
administered with the pharmaceutical composition; having chronic
lung disease, chronic asthma; obesity with body mass index
[BMI]>35, having symptoms of COVID-19 selected from fever,
cough, sore throat, malaise, headache, muscle pain, nausea,
vomiting, diarrhea, loss of taste and smell, or a combination
thereof, having shortness of breath, dyspnea, or abnormal chest
imaging, having evidence of lower respiratory disease during
clinical assessment or imaging, having saturation of oxygen (SpO2)
.gtoreq.94% on room air at sea level, having severe symptoms of the
infection of the SARS-CoV-2, having SpO2 <94% on room air at sea
level, having a ratio of arterial partial pressure of oxygen to
fraction of inspired oxygen (PaO2/FiO2) <300 mmHg, respiratory
frequency>30 breaths per minute, lung infiltrates >50%,
having active symptoms of antibody-dependent enhancement (ADE),
having a history of antibody-dependent enhancement (ADE), being
allergic to an antibody treatment, being a hospital inpatient
requiring supportive management of complications of severe
infection of the SARS-CoV-2 selected from pneumonia, hypoxemic
respiratory failure/ARDS, sepsis and septic shock, cardiomyopathy
and arrhythmia, acute kidney injury, and complications from
prolonged hospitalization including secondary bacterial and fungal
infections, thromboembolism, gastrointestinal bleeding, critical
illness polyneuropathy/myopathy, or a combination thereof.
[0715] In some embodiments, the pharmaceutical composition can
further comprise one or more bioactive agent that can comprise a
therapeutic agent or a prophylactic agent selected from an
anti-viral agent, an antiviral peptide, an anti-viral antibody, an
anti-viral compound, an anti-viral cytokine, an anti-viral
oligonucleotide, an RNA dependent RNA polymerase inhibitor, a
non-nucleoside reverse transcriptase inhibitor (NNRTI), nucleoside
reverse transcriptase inhibitor (NRTI), purine nucleoside,
antiviral interferon, adamantine antiviral compound, remdesivir,
chloroquine, hydroxychloroquine, lopinavir, ritonavir, APN01,
favilavir, mesalazine, toremifene, eplerenone, paroxetine,
sirolimus, dactinomycin, irbesartan, emodin, mercaptopurine,
melatonin, quinacrine, carvedilol, colchicine, camphor, equilin,
oxymetholone, nafamosta, camostat, baricitinib, darunavir,
ribavirin, galidesivir, BCX-4430, Arbidol, nitazoxanide, one or
more derivatives thereof, or any combination thereof. In some
cases, the pharmaceutical composition can further comprise
infliximab, abalizumab, ustekinumab, immunomodulators such as
methotrexate, 6MP, azathioprine, or a combination thereof.
chronic
[0716] Methods of Treatment or Prevention
[0717] The present disclosure also provides methods of treating
SARs-CoV-2 infection or a disease, disorder or condition associated
with SARs-CoV-2 infection in a subject, comprising administering to
the subject a therapeutically effective amount of one or more of
the antibody or antigen-binding fragment thereof provided herein,
or one or more polynucleotides encoding one or more of the antibody
or antigen-binding fragment thereof provided herein, or the
pharmaceutical composition provided herein.
[0718] In certain embodiments, the therapeutically effective amount
can be an amount effective to decrease SARs-COV-2 titers, or to
alleviate one or more disease symptoms, viremia, or any other
measurable manifestation of SARS-CoV-2 infection in the treated
subject or population, whether by inducing the regression of or
inhibiting the progression of symptom(s) associated with SARs-COV-2
infection by any clinically measurable degree. Decrease in
SARs-COV-2 titers can be measured in the lung, for example, by the
concentration of SARs-COV-2 in sputum samples or a lavage from the
lungs from a mammal. Alleviation of a disease symptom can be
assessed by any clinical measurement typically used by physicians
or other skilled healthcare providers to assess the severity or
progression status of that symptom. Exemplary symptoms associated
with SARs-COV-2 infection include, without limitation, fever, dry
cough, shortness in breath, pain or pressure in the chest, new
confusion or inability to arouse, bluish lips or face, loss of
sense of smell and/or loss of sense of taste.
[0719] A subject in need of treatment include, for example, those
already infected with SARS-CoV-2 (symptomatic or asymptomatic) or
inflicted with a condition resulting from infection of SARS-CoV-2.
Subjects partially or totally recovered from infection of
SARS-CoV-2 might also be in need of treatment. In certain
embodiments, the subject is human.
[0720] The present disclosure also provides methods of preventing
SARs-CoV-2 infection, or a disease, disorder or condition
associated with SARs-COV-2 infection in a subject, comprising
administering to the subject a prophylactically effective amount of
one or more of the antibody or antigen-binding fragment thereof
provided herein, or one or more polynucleotides encoding one or
more of the antibody or antigen-binding fragment thereof provided
herein, or the pharmaceutical composition provided herein.
Prevention encompasses inhibiting or reducing the spread of
SARS-CoV-2 or inhibiting or reducing the onset, development or
progression of one or more of the symptoms associated with
infection with SARS-CoV-2.
[0721] In certain embodiments, the prophylactically effective
amount can be an amount effective to neutralize SARs-COV-2 in the
respiratory tract, lungs and/or other affected areas such as eyes,
noses and mouth, in order block infection, or effective to
ameliorate at least one symptom associated with SARs-COV-2
infection. Whether a symptom has been ameliorated can be assessed
by any clinical measurement typically used by physicians or other
skilled healthcare providers to assess the severity or progression
status of that symptom or in certain instances will ameliorate the
need for hospitalization.
[0722] A subject in need of prevention include, for example, those
in which infection with SARS-CoV-2 is to be prevented, or those who
are at risk for SARS-CoV-2 infection. In certain embodiments, the
subject is human.
[0723] The term "disease, disorder or condition associated with
SARS-COV-2 infection" as used herein include those that are caused
by or related to SARs-COV-2 infection, such as, upper or lower
respiratory tract infections, pharyngitis, pneumonia,
tracheobronchitis, bronchiolitis, bronchitis, acute respiratory
distress syndrome, diarrhea, and any related infections or
inflammatory disorders.
[0724] The methods of treatment or prevention provided herein are
also suitable for gene therapy by transfer of polynucleotide
sequences encoding the antibody product or fragment thereof in a
subject, such that the polynucleotide can be expressed in the
subject to produce the antibody in vivo. The polynucleotide
provided herein can be administered to a subject by, for example,
transfection techniques such as electroporation and hydrodynamic
injection, which are suitable for administration of naked
polynucleotides. For polynucleotides in the form of viral vectors
such as AAV, it can be administered via local injection (e.g.
intramuscular, intranasal, intradermal, subcutaneous, etc.) or
systematic administration (e.g. intravenous administration).
[0725] In certain embodiments, the methods can comprise
administering to the subject a therapeutically effective amount or
a prophylactically effective amount of a combination of two or more
of the antibodies (or the antigen-binding fragment thereof)
provided herein. In certain embodiments, the two or more antibodies
comprises a first antibody comprising heavy chain CDR sequences and
light chain CDR sequences derived from P2C-1F11, and a second
antibody comprising heavy chain CDR sequences and light chain CDR
sequences derived from antibody P2B-2F6. In certain embodiments,
the two or more antibodies or the antigen binding fragments thereof
bind to different epitopes in RBD of spike protein of SARS-CoV-2.
In certain embodiments, the two or more antibodies comprise a first
antibody comprising P2C-1F11, and a second antibody which is
selected from the group consisting of P2C-1A3, P2C-1C10, P2B-2F6,
P2B-1G5, and P2A-1B3. In certain embodiments, the two or more
antibodies comprise a first antibody comprising P2C-1A3 and a
second antibody which is selected from the group consisting of
P5A-3C8, P5A-1D2, P22A-1D1, P2C-1F11, and P2A-1B3, or an antigen
binding fragment thereof. In certain embodiments, the two or more
antibodies comprise a first antibody comprising P2B-2F6 and a
second antibody which is selected from the group consisting of
P2C-1C10, P2C-1F11, P2B-1G5, and P2A-1B3, or an antigen binding
fragment thereof. In certain embodiments, the two or more
antibodies comprises a first antibody comprising P2A-1B3 and a
second antibody which selected from the group consisting of
P5A-3C8, P5A-1D2, P22A-1D1, P2C-1A3, P2C-1C10, P2C-1F11, P2B-2F6,
and P2A-1A10, or an antigen binding fragment thereof. In some
embodiments, the two or more antibodies comprise a first antibody
which comprises P2C-1C10 or an antigen binding fragment thereof,
and a second antibody selected from the group consisting of
P5A-3C8, P5A-1D2, P22A-1D1, P2C-1A3, P2C-1F11, and P2A-1B3, or an
antigen binding fragment thereof.
[0726] The antibodies or antigen-binding fragments thereof provided
herein may be administered by any route known in the art, such as
for example parenteral (e.g. subcutaneous, intraperitoneal,
intravenous, including intravenous infusion, intramuscular, or
intradermal injection) or non-parenteral (e.g. oral, intranasal,
intraocular, sublingual, rectal, or topical) routes.
[0727] In some embodiments, this disclosure is directed to a method
for treating or preventing a disease in a subject in need thereof,
the method can comprise administering an effective dosage of any
one of the pharmaceutical compositions disclosed herein to the
subject;
[0728] wherein the pharmaceutical composition can be configured to
be administered to the subject to maintain a plasma concentration
of the modified antibody or an antigen-binding fragment thereof in
a therapeutic effective range of from 10 .mu.g/mL to 3500 .mu.g/mL
for a time period in a range of from 1 day to 12 months after
administering the pharmaceutical composition; and
[0729] wherein the subject can be infected with, exhibiting one or
more symptoms of being infected with, or at risk of being infected
with the SARS-CoV-2.
[0730] The method disclosed herein can be used for preventing
infection of the SARS-CoV-2 in a subject who is at risk of being
infected, such as a healthy person who may get in contact with
another person who has or had the SARS-CoV-2 infection with or
without symptoms, a person who provides case to or handles
materials related from another person who has or had the SARS-CoV-2
infection with or without symptoms, such as a healthcare personnel,
an emergency responder, a medical diagnosis service personnel, a
senior home service provider, or a combination thereof.
[0731] In some embodiments, the pharmaceutical composition can be
administered to the subject having no symptoms or free from known
infections of the SARS-CoV-2, prior to the subject being infected
with the SARS-CoV-2, prior to the subject exhibiting any symptoms
of the infection of the SARS-CoV-2, or a combination thereof.
[0732] In some embodiments, the pharmaceutical composition can be
configured to be administered to the subject to maintain the plasma
concentration of the modified antibody or an antigen-binding
fragment thereof in a therapeutic effective range of from 10
.mu.g/mL to 1500 .mu.g/mL for a time period ranging from 3 to 12
months after the administration and wherein the administration is a
single administration. In some embodiments, the pharmaceutical
composition can be configured to be administered to the subject to
maintain the plasma concentration of the modified antibody or an
antigen-binding fragment thereof in a therapeutic effective range
of from 10 .mu.g/mL to 1500 .mu.g/mL, 20 .mu.g/mL to 1500 .mu.g/mL,
30 .mu.g/mL to 1500 .mu.g/mL, 40 .mu.g/mL to 1500 .mu.g/mL, 50
.mu.g/mL to 1500 .mu.g/mL, 60 .mu.g/mL to 1500 .mu.g/mL, 70
.mu.g/mL to 1500 .mu.g/mL, 80 .mu.g/mL to 1500 .mu.g/mL, 90
.mu.g/mL to 1500 .mu.g/mL, 100 .mu.g/mL to 1500 .mu.g/mL, 150
.mu.g/mL to 1500 .mu.g/mL, 200 .mu.g/mL to 1500 .mu.g/mL, 300
.mu.g/mL to 1500 .mu.g/mL, 400 .mu.g/mL to 1500 .mu.g/mL, 500
.mu.g/mL to 1500 .mu.g/mL, 600 .mu.g/mL to 1500 .mu.g/mL, 700
.mu.g/mL to 1500 .mu.g/mL, 800 .mu.g/mL to 1500 .mu.g/mL, 900
.mu.g/mL to 1500 .mu.g/mL, 1000 .mu.g/mL to 1500 .mu.g/mL, 1100
.mu.g/mL to 1500 .mu.g/mL, 1200 .mu.g/mL to 1500 .mu.g/mL, 1300
.mu.g/mL to 1500 .mu.g/mL or 1400 .mu.g/mL to 1500 .mu.g/mL,
wherein the disclosure of these ranges is intended as a continuous
range including every value between the minimum and maximum values.
In some embodiments, the time period can range from 3 to 12 months,
4 to 12 months, 5 to 12 months, 6 to 12 months, 7 to 12 months, 8
to 12 months, 9 to 12 months, 10 to 12 months or 11 to 12 months,
after administration of the pharmaceutical composition and wherein
the administration can be a single administration. The plasma
concentration of the modified antibody or an antigen-binding
fragment thereof to reach the above mentioned range within a day
and can maintain within the above mentioned range for the indicated
time periods disclosed above.
[0733] In some embodiments, the subject can be a person having no
symptoms or free from known infections of the SARS-CoV-2, or
treatment of a patient being a symptomatic non-hospitalized adult
with COVID-19 caused by SARS-CoV-2 infection, aged 60 years and
older, any age having at least one of the following conditions
selected from smoking, exogenous or endogenous immunosuppression
having HIV infection with CD4 count <200 cells/mm3, receiving
corticosteroids equivalent to prednisone .gtoreq.20 mg daily for at
least 14 consecutive days within 30 days prior to be administered
with the pharmaceutical composition, receiving one or more
biologics therapeutical agents, one or more immunomodulators,
cancer chemotherapy within 90 days prior to be administered with
the pharmaceutical composition; having chronic lung disease,
chronic asthma; obesity with body mass index [BMI]>35, having
symptoms of COVID-19 selected from fever, cough, sore throat,
malaise, headache, muscle pain, nausea, vomiting, diarrhea, loss of
taste and smell, or a combination thereof, having shortness of
breath, dyspnea, or abnormal chest imaging, having evidence of
lower respiratory disease during clinical assessment or imaging,
having saturation of oxygen (SpO2) .gtoreq.94% on room air at sea
level, having severe symptoms of the infection of the SARS-CoV-2,
having SpO2 <94% on room air at sea level, having a ratio of
arterial partial pressure of oxygen to fraction of inspired oxygen
(PaO2/FiO2) <300 mmHg, respiratory frequency>30 breaths per
minute, lung infiltrates >50%, having active symptoms of
antibody-dependent enhancement (ADE), having a history of
antibody-dependent enhancement (ADE), being allergic to an antibody
treatment, being a hospital inpatient requiring supportive
management of complications of severe infection of the SARS-CoV-2
selected from pneumonia, hypoxemic respiratory failure/ARDS, sepsis
and septic shock, cardiomyopathy and arrhythmia, acute kidney
injury, and complications from prolonged hospitalization including
secondary bacterial and fungal infections, thromboembolism,
gastrointestinal bleeding, critical illness
polyneuropathy/myopathy, or a combination thereof.
[0734] In some embodiments, the subject can be a person 60 years
and older, 65 years and older, 70 years and older, 75 years and
older, 80 years and older, 85 years and older or 90 years and
older.
[0735] In some embodiments of the method disclosed herein, the
pharmaceutical composition can be configured to be administered to
the subject to maintain the plasma concentration of the modified
antibody or an antigen-binding fragment thereof in a therapeutic
effective range of from 30 .mu.g/mL to 3500 .mu.g/mL for a time
period ranging from 1 to 4 weeks after the administration and
wherein the administration is a single administration. In some
embodiments, the pharmaceutical composition can be configured to be
administered to the subject to maintain the plasma concentration of
the modified antibody or an antigen-binding fragment thereof in a
therapeutic effective range of from 10 .mu.g/mL to 3500 .mu.g/mL,
20 .mu.g/mL to 3500 .mu.g/mL, 30 .mu.g/mL to 3500 .mu.g/mL, 40
.mu.g/mL to 3500 .mu.g/mL, 50 .mu.g/mL to 3500 .mu.g/mL, 60
.mu.g/mL to 3500 .mu.g/mL, 70 .mu.g/mL to 3500 .mu.g/mL, 80
.mu.g/mL to 3500 .mu.g/mL, 90 .mu.g/mL to 3500 .mu.g/mL, 100
.mu.g/mL to 3500 .mu.g/mL, 150 .mu.g/mL to 3500 .mu.g/mL, 200
.mu.g/mL to 3500 .mu.g/mL, 300 .mu.g/mL to 3500 .mu.g/mL, 400
.mu.g/mL to 3500 .mu.g/mL, 500 .mu.g/mL to 3500 .mu.g/mL, 600
.mu.g/mL to 3500 .mu.g/mL, 700 .mu.g/mL to 3500 .mu.g/mL, 800
.mu.g/mL to 3500 .mu.g/mL, 900 .mu.g/mL to 3500 .mu.g/mL, 1000
.mu.g/mL to 3500 .mu.g/mL, 1100 .mu.g/mL to 3500 .mu.g/mL, 1200
.mu.g/mL to 3500 .mu.g/mL, 1300 .mu.g/mL to 3500 .mu.g/mL, 1400
.mu.g/mL to 3500 .mu.g/mL, 1500 .mu.g/mL to 3500 .mu.g/mL, 1600
.mu.g/mL to 3500 .mu.g/mL, 1700 .mu.g/mL to 3500 .mu.g/mL, 1800
.mu.g/mL to 3500 .mu.g/mL, 1900 .mu.g/mL to 3500 .mu.g/mL, 2000
.mu.g/mL to 3500 .mu.g/mL, 2100 .mu.g/mL to 3500 .mu.g/mL, 2200
.mu.g/mL to 3500 .mu.g/mL, 2300 .mu.g/mL to 3500 .mu.g/mL, 2400
.mu.g/mL to 3500 .mu.g/mL, 2500 .mu.g/mL to 3500 .mu.g/mL, 2600
.mu.g/mL to 3500 .mu.g/mL, 2700 .mu.g/mL to 3500 .mu.g/mL, 2800
.mu.g/mL to 3500 .mu.g/mL, 2900 .mu.g/mL to 3500 .mu.g/mL, 3000
.mu.g/mL to 3500 .mu.g/mL, 3100 .mu.g/mL to 3500 .mu.g/mL, 3200
.mu.g/mL to 3500 .mu.g/mL, 3300 .mu.g/mL to 3500 .mu.g/mL or 3400
.mu.g/mL to 3500 .mu.g/mL, wherein the disclosure of these ranges
is intended as a continuous range including every value between the
minimum and maximum values. In some embodiments, the time period
can range from 1 to 4 weeks, 2 to 4 weeks or 3 to 4 weeks after
administration of the pharmaceutical composition and wherein the
administration can be a single administration. The plasma
concentration of the modified antibody or an antigen-binding
fragment thereof to reach the above mentioned range within a day
and can maintain within the above mentioned range for the indicated
time periods disclosed above.
[0736] In some embodiments, the pharmaceutical composition can be
administered to maintain a high plasma concentration, such as 30
.mu.g/mL to 3500 .mu.g/mL, of the modified antibody or an
antigen-binding fragment thereof immediately after administration,
such as 1 day to a few days or 1 to 4 weeks, for treating a patient
with the disease or symptoms of the infection of the SARS-CoV-2. In
some embodiments, the pharmaceutical composition can be
administered to maintain a desired plasma concentration, such as 10
.mu.g/mL to 1500 .mu.g/mL, of the modified antibody or an
antigen-binding fragment thereof and maintain within the desired
range for 3 to 12 months, for preventing a person from being
infected with the SARS-CoV-2. As used herein the "plasma
concentration" or "serum concentration" may be used interchangeably
for the concentration of the modified antibody or an
antigen-binding fragment thereof in the blood of a patient.
[0737] In some embodiments, plasma concentration of the modified
antibody or an antigen-binding fragment thereof can be at about
100-300 times of in vitro IC.sub.90 for at least 3 to 6 weeks for
treating a patient with the SARS-CoV-2 infection or symptoms of the
SARS-CoV-2 infection. In some embodiments, plasma concentration of
the modified antibody or an antigen-binding fragment thereof can be
at about 10-50 times of in vitro IC.sub.90 for at least 6-month for
preventing the SARS-CoV-2 infection or symptoms of the SARS-CoV-2
infection.
[0738] In some embodiments of the method disclosed herein, the
modified antibody or the antigen-binding fragment thereof can be
configured to have a half-life (T.sub.1/2) in a range of from 50 to
120 days in the subject. In some embodiments, the half-life
(T.sub.1/2) can be in a range of from 50 to 120 days, 60 to 120
days, 70 to 120 days, 80 to 120 days, 90 to 120 days, 100 to 120
days or 110 to 120 days, in the subject.
[0739] In some embodiments of the method disclosed herein, the
pharmaceutical composition can be configured to be administered to
the subject in a range of from 150 mg/m.sup.2 to 5000 mg/m.sup.2.
In some cases, the pharmaceutical composition can be administered
to the subject in a dosage range of from 150 to 5000 mg/m.sup.2,
200 to 5000 mg/m.sup.2, 300 to 5000 mg/m.sup.2, 400 to 5000
mg/m.sup.2, 500 to 5000 mg/m.sup.2, 600 to 5000 mg/m.sup.2, 700 to
5000 mg/m.sup.2, 800 to 5000 mg/m.sup.2, 900 to 5000 mg/m.sup.2,
1000 to 5000 mg/m.sup.2, 1200 to 5000 mg/m.sup.2, 1400 to 5000
mg/m.sup.2, 1600 to 5000 mg/m.sup.2, 1800 to 5000 mg/m.sup.2, 2000
to 5000 mg/m.sup.2, 2200 to 5000 mg/m.sup.2, 2400 to 5000
mg/m.sup.2, 2600 to 5000 mg/m.sup.2, 2800 to 5000 mg/m.sup.2, 3000
to 5000 mg/m.sup.2, 3200 to 5000 mg/m.sup.2, 3400 to 5000
mg/m.sup.2, 3600 to 5000 mg/m.sup.2, 3800 to 5000 mg/m.sup.2, 4000
to 5000 mg/m.sup.2, 4200 to 5000 mg/m.sup.2, 4400 to 5000
mg/m.sup.2, 4600 to 5000 mg/m.sup.2 or 4800 to 5000 mg/m.sup.2,
wherein the disclosure of these ranges is intended as a continuous
range including every value between the minimum and maximum
values.
[0740] In some embodiments of the method disclosed herein, the
pharmaceutical composition can be configured to be administered to
the subject in a range of from 300 mg to 8000 mg. In some cases,
the pharmaceutical composition can be administered to the subject
in a range of from 300 to 8000 mg, 400 to 8000 mg, 500 to 8000 mg,
600 to 8000 mg, 700 to 8000 mg, 800 to 8000 mg, 900 to 8000 mg,
1000 to 8000 mg, 1200 to 8000 mg, 1400 to 8000 mg, 1600 to 8000 mg,
1800 to 8000 mg, 2000 to 8000 mg, 2500 to 8000 mg, 3000 to 8000 mg,
3500 to 8000 mg, 4000 to 8000 mg, 4500 to 8000 mg, 5000 to 8000 mg,
5500 to 8000 mg, 6000 to 8000 mg, 6500 to 8000 mg, 7000 to 8000 mg
or 7500 to 8000 mg. In some cases, the pharmaceutical composition
can be administered to the subject in a range of from 5 to 150
mg/kg, 10 to 150 mg/kg, 15 to 150 mg/kg, 20 to 150 mg/kg, 25 to 150
mg/kg, 30 to 150 mg/kg, 35 to 150 mg/kg, 40 to 150 mg/kg, 45 to 150
mg/kg, 50 to 150 mg/kg, 55 to 150 mg/kg, 60 to 150 mg/kg, 65 to 150
mg/kg, 70 to 150 mg/kg, 75 to 150 mg/kg, 80 to 150 mg/kg, 85 to 150
mg/kg, 90 to 150 mg/kg, 95 to 150 mg/kg, 100 to 150 mg/kg, 110 to
150 mg/kg, 120 to 150 mg/kg, 130 to 150 mg/kg or 140 to 150 mg/kg,
of the body weight of the subject.
[0741] In some embodiments, the pharmaceutical composition can be
configured to have the modified antibody at a concentration in a
range of from 10 mg/mL to 150 mg/mL. In some cases, the
pharmaceutical composition can be configured to have the modified
antibody at a concentration at 10 mg/mL to 150 mg/mL, 20 mg/mL to
150 mg/mL, 30 mg/mL to 150 mg/mL, 40 mg/mL to 150 mg/mL, 50 mg/mL
to 150 mg/mL, 60 mg/mL to 150 mg/mL, 70 mg/mL to 150 mg/mL, 80
mg/mL to 150 mg/mL, 90 mg/mL to 150 mg/mL, 100 mg/mL to 150 mg/mL,
110 mg/mL to 150 mg/mL, 120 mg/mL to 150 mg/mL, 130 mg/mL to 150
mg/mL or 140 mg/mL to 150 mg/mL. The concentration can be the total
protein concentration of the antibody in the pharmaceutical
composition.
[0742] In some cases of the method disclosed herein, the
pharmaceutical composition can be administered to the subject via
intravenous injection (IV), intramuscular injection (IM),
subcutaneous (SC) injection, or a combination thereof.
[0743] In some embodiments of the method disclosed herein, the
effective dosage can be determined by a dosing process that can
comprise determining concentration progression data based on
calculated or measured pharmacokinetics (PK), testing plasma
concentrations over a testing period of time, predicted plasma
concentrations over a prediction period of time, or a combination
thereof, of the modified antibody or the antigen-binding fragment
thereof, and producing the effective dosage based on the
concentration progression data. In some embodiments, the effective
dosage can be determined by predicted plasma concentrations over a
prediction period of time, wherein the predicted plasma
concentrations can be produced by measuring actual plasma
concentrations of the modified antibody in a subject selected form
a primate or a human over a measurement period of time to produce
measured concentration data and interpolating and extrapolating the
measured concentration data to produce the predicted plasma
concentrations in a selected prediction period of time.
[0744] In some embodiments of the method disclosed herein, the
effective dosage can be selected to maintain the plasma
concentration in a range of from 10 .mu.g/mL to 1500 .mu.g/mL in 3
to 12 months after the administration. Such effective dosage can be
suitable for preventing the disease in a subject for an extended
period of time, such as 3 to 12 months.
[0745] In some embodiments of the method disclosed herein, the
effective dosage can be selected to maintain the plasma
concentration in a range of from 1500 .mu.g/mL to 3500 .mu.g/mL in
1 day to 2 months after the administration. Such high effective
dosage can be suitable for treating the disease for a shorter
period of time, for example, from 1 day to 60 days.
[0746] In some embodiments of the method disclosed herein, the
pharmaceutical composition further comprises one or more subsequent
modified antibodies selected from a first subsequent modified
antibody comprising two antigen-binding domains each having same or
different affinities to the SARS-CoV-2, a second subsequent
modified antibody comprising a first antigen-binding domain having
a binding affinity to the SARS-CoV-2 and a second antigen-binding
domain having a binding affinity to a second pathogen that is
different from the SARS-CoV-2, a third subsequent modified antibody
comprising two antigen-binding domains each having a same or
different binding affinity to the second pathogen, or a combination
thereof. As mentioned above, the term "different affinities to the
SARS-CoV-2" refers affinity that can bind to a different epitope or
binding site of the SARS-CoV-2, a different affinity level that can
bind to the same epitope or binding site of the SARS-CoV-2, or a
combination thereof. The binding affinity to the second pathogen
can be selected from a binding affinity to SARS-CoV, MERS-CoV, one
or more bacteria, one or more fungus, one or more viruses, one or
more parasites, a part thereof, or a combination thereof.
[0747] In some embodiments of the method disclosed herein can
further comprise administering a pharmaceutically effective amount
of one or more bioactive agents to the subject simultaneously or
sequentially with the pharmaceutical composition, wherein the
bioactive agent comprises a therapeutic agent or a prophylactic
agent selected from an anti-viral agent, an antiviral peptide, an
anti-viral antibody, an anti-viral compound, an anti-viral
cytokine, an anti-viral oligonucleotide, an RNA dependent RNA
polymerase inhibitor, a non-nucleoside reverse transcriptase
inhibitor (NNRTI), nucleoside reverse transcriptase inhibitor
(NRTI), purine nucleoside, antiviral interferon, adamantine
antiviral compound, remdesivir, chloroquine, hydroxychloroquine,
lopinavir, ritonavir, APN01, favilavir, mesalazine, toremifene,
eplerenone, paroxetine, sirolimus, dactinomycin, irbesartan,
emodin, mercaptopurine, melatonin, quinacrine, carvedilol,
colchicine, camphor, equilin, oxymetholone, nafamosta, camostat,
baricitinib, darunavir, ribavirin, galidesivir, BCX-4430, Arbidol,
nitazoxanide, one or more derivatives thereof, or any combination
thereof.
[0748] In some embodiments, the antibodies or antigen-binding
fragments thereof provided herein may be administered alone or in
combination a therapeutically effective amount of a second
bioactive agent. The second bioactive agent can be a therapeutic
agent or a prophylactic agent.
[0749] In some embodiments, the second therapeutic agent is an
anti-viral agent. In some embodiments, the anti-viral agent
comprises an antiviral peptide, an anti-viral antibody, an
anti-viral compound, an anti-viral cytokine, or an anti-viral
oligonucleotide. In some embodiments, the anti-viral agent is an
RNA dependent RNA polymerase inhibitor, a non-nucleoside reverse
transcriptase inhibitor (NNRTI), nucleoside reverse transcriptase
inhibitor (NRTI), purine nucleoside, antiviral cytokines such as
interferon, adamantine antiviral compound, anti-RBD antibody,
anti-S1 antibody, anti-S2 antibody, siRNAs Targeting mRNA of
coronavirus proteins M, N, or E (Chinese patent applications
CN101173275 and CN1648249), siRNAs targeting replicase and RNA
polymerase region (US patent application US20050004063), RNA
Aptamers (Korean patent applications KR2009128837 and KR
2012139512), ribozymes (Japanese patent application JP2007043942),
antisense oligonucleotides (PCT patent application WO2005023083),
or any other suitable antiviral agent. In certain embodiments, the
anti-viral compound is selected from the group consisting of
remdesivir, chloroquine, hydroxychloroquine, lopinavir, ritonavir,
APN01, favilavir, mesalazine, toremifene, eplerenone, paroxetine,
sirolimus, dactinomycin, irbesartan, emodin, mercaptopurine,
melatonin, quinacrine, carvedilol, colchicine, camphor, equilin,
oxymetholone, nafamosta, camostat, baricitinib, darunavir,
ribavirin, galidesivir, BCX-4430, Arbidol, nitazoxanide,
derivatives thereof, or any combination thereof. More examples of
potentially useful anti-viral agents for SARS-CoV-2 reviewed by C.
Liu et al, ACS Cent. Sci. 2020, 6, 3, 315-331, which is incorporate
herein to its entirety.
[0750] In certain embodiments, the second bioactive agent (e.g.
prophylactic agent) can be a SARS-CoV-2 vaccine (e.g. mRNA-1273 by
Moderna, an AAV-based vaccine capable of expressing an SARS-CoV-2
immunogen), an antibody (e.g. directed to SARS-CoV-2), lymphokines,
hematopoietic growth factors (such as IL-2, IL-3, IL-7, and IL-15),
which can for example serve to increase the number or activity of
effector cells which interact with the antibodies.
[0751] In certain embodiments, the second bioactive agent can
comprise hormonal therapy, immunotherapy, and anti-inflammatory
agents.
[0752] In certain of these embodiments, an antibody or
antigen-binding fragment thereof provided herein may be
administered simultaneously with the one or more additional
bioactive agents, and in certain of these embodiments the antibody
or antigen-binding fragment thereof and the additional therapeutic
agent(s) may be administered as part of the same pharmaceutical
composition. However, an antibody or antigen-binding fragment
thereof administered "in combination" with another bioactive agent
does not have to be administered simultaneously with or in the same
composition as the agent. An antibody or antigen-binding fragment
thereof administered prior to or after another agent is considered
to be administered "in combination" with that agent as the phrase
is used herein, even if the antibody or antigen-binding fragment
and the second agent are administered via different routes. Where
possible, additional bioactive agents administered in combination
with the antibodies or antigen-binding fragments thereof disclosed
herein are administered according to the schedule listed in the
product information sheet of the additional therapeutic agent, or
according to the Physicians' Desk Reference 2003 (Physicians' Desk
Reference, 57th Ed; Medical Economics Company; ISBN: 1563634457;
57th edition (November 2002)) or protocols well known in the
art.
[0753] One advantage of the modified antibody, the pharmaceutical
composition and the method disclosed herein is that the modified
antibody or an antigen-binding fragment thereof comprising at least
an antigen-binding domain having an antigen-binding affinity and a
covalently linked modified human IgG constant domain, wherein the
modified human IgG constant domain comprises a substitution with
tyrosine at amino acid residue 252, a substitution with threonine
at amino acid residue 254, and a substitution with glutamic acid at
amino acid residue 256, numbered according to the EU index as in
Kabat, said modified antibody has an increased affinity for FcRn
compared to the affinity to FcRn of an antibody having a wild type
human IgG constant domain. Such antibody can have extended
half-life in vivo. Not wishing to be bound by a particular theory
or a mechanism, Applicants believe that the increased affinity to
FcRn can help the antibody to escape intracellular degradations and
increase the antibody recycling, therefore increasing the amount of
the antibody remaining in the blood stream of the subject
preventing or treating the disease.
[0754] Another advantage of the modified antibody, the
pharmaceutical composition and the method disclosed herein is that
the modified antibody can have a reduced affinity to human
Fc.gamma. receptors (Fc.gamma.R) that belong to the immunoglobulin
superfamily. The reduced affinity to human Fc.gamma.R can help to
reduce certain immune response side effects, such as
antibody-dependent enhancement (ADE).
[0755] Methods of Virus Detection
[0756] In another aspect, the present disclosure provides a method
of detecting presence or amount of SARS-CoV-2 virus antigen in a
sample. In some embodiments, the SARS-CoV-2 virus antigen comprises
spike protein, or comprises the SARS-CoV-2 virus particle. In some
embodiments, the method comprises contacting the sample with the
antibody or antigen binding fragment disclosed herein, and
determining the presence or the amount of the SARS-CoV-2 virus
antigen in the sample.
[0757] In certain embodiments, the anti-SARS-CoV-2 antibody
disclosed herein is used in a method of diagnosing a subject
suffering from a disorder (e.g., SARS-CoV-2 infection), the method
comprising: determining the presence or amount of SARS-CoV-2 virus
antigen in a sample obtained from the subject by contacting the
sample with an anti-SARS-CoV-2 antibody of the disclosure and
detecting the presence of the bound antibody.
[0758] Any sample suspected of containing SARS-CoV-2 virus can be
used. In some embodiments, a suitable sample can be obtained from
respiratory tract of the subject, for example, an upper respiratory
nasopharyngeal swab (NP), oropharyngeal swabs (OP), sputum, a lower
respiratory tract aspirate, bronchoalveolar lavage sample,
nasopharyngeal wash, nasopharyngeal aspirate, nasal aspirate, a
nasal swap, a throat swap, a bronchoalveolar lavage fluid (BALF), a
cell or tissue sample from respiratory tract or from lung, and the
like. In some embodiments, a suitable sample can be a body fluid
sample such as a whole blood sample, a serum sample, or a plasma
sample. In some embodiments, a suitable sample can be a urine
sample or a stool sample.
[0759] The presence or level of SARS-CoV-2 virus antigen in a
sample can be determined based on the detection of the presence or
level of the complex of the virus antigen bound by the antibody or
the antigen binding fragment thereof disclosed herein. Any suitable
methods can be used for such detection, for example, by
immunoassays such as immunohistochemistry (IHC), immunofluorescence
(IF), immunoblotting (e.g., Western blotting), flow cytometry
(e.g., FACS.TM.), Enzyme-linked Immunosorbant Assay (ELISA), enzyme
immunoassay (ETA), and radioimmunoassay (RIA).
[0760] For a review of immunological and immunoassay procedures,
see Basic and Clinical Immunology (Stites & Terr eds., 7.sup.th
ed. 1991). Moreover, the immunoassays can be performed in any of
several configurations, which are reviewed extensively in Enzyme
Immunoassay (Maggio, ed., 1980); and Harlow & Lane, supra. For
a review of the general immunoassays, see also Methods in Cell
Biology: Antibodies in Cell Biology, volume 37 (Asai, ed. 1993);
Basic and Clinical Immunology (Stites & Terr, eds., 7.sup.th
ed. 1991).
[0761] In certain embodiments, the antibodies or the antigen
binding fragments thereof disclosed herein are detectably labeled,
or are not labeled but can react with a second molecule which is
detectably labeled (e.g. a detectably labeled secondary
antibody).
[0762] In certain embodiments, the antibodies or the antigen
binding fragments thereof disclosed herein may be immobilized on a
solid substrate. The immobilization can be via covalent linking or
non-covalent attachment (e.g. coating). Examples of solid substrate
include porous and non-porous materials, latex particles, magnetic
particles, microparticles, strips, beads, membranes, microtiter
wells and plastic tubes. The choice of solid phase material and
method of detectably labeling can be determined based upon desired
assay format performance characteristics.
[0763] The level of the SARS-CoV-2 antigen can be determined, for
example, by normalizing to a control value or to a standard curve.
The control value can be predetermined, or determined
concurrently.
[0764] The assays and methods provided herein for the measurement
of the level of the SARS-CoV-2 antigen can be adapted or optimized
for use in automated and semi-automated systems, or point of care
assay systems.
[0765] Methods of Antibody Detection
[0766] In another aspect, the present disclosure provides a method
of detecting presence or amount of an antibody capable of
specifically binding to RBD of the spike protein of SARS-CoV-2 in a
sample, comprising contacting the sample with a polypeptide
comprising an amino acid sequence comprising SEQ ID NO: 128, and
determining the presence or the level of the antibody in the
sample. In some embodiments, the absence of the antibody in the
sample or the level of the antibody in the sample being below a
threshold indicates that the subject is more likely to suffer from
disease progression.
[0767] In another aspect, the present disclosure provides a method
of determining or predicting the likelihood of disease progression
in a subject infected with SARS-CoV-2, the method comprising:
contacting a sample obtained from the subject with a polypeptide
comprising an amino acid sequence comprising SEQ ID NO: 128, and
detecting the presence or the level of an antibody in the sample
wherein the antibody is capable of specifically binding to RBD of
the spike protein of the SARS-CoV-2, wherein the subject is likely
to experience disease progression when the antibody in the sample
is absent or is below a threshold.
[0768] A subject infected with SARS-CoV-2 can produce antibodies
against the SARS-CoV-2 antigens. Such antibodies produced by human
immune system are polyclonal, and can bind to different antigens or
epitopes of SARS-CoV-2. Without wishing to be bound by any theory,
it is unexpectedly found by the inventors that the presence or
level of the antibodies specific to the RBD of the spike protein of
the SARS-CoV-2 can be indicative of likelihood of disease
progression in the subject. Antibodies capable of specifically
binding to the RBD of the spike protein of the SARS-CoV-2
("RBD-specific antibodies") are found by the inventors to be
capable of effectively competing with ACE2 receptor for binding to
the RBD, and also provide for SARS-CoV-2 virus neutralizing
activity. The presence of such a RBD-specific antibody can be
associated with an effective immune response to the SARS-CoV-2, and
the titer of such RBD-specific antibody in the body may correlate
to the prognosis of the SARS-CoV-2 infection or a disease, disorder
or condition associated with SARs-CoV-2 infection.
[0769] A threshold of the level of the RBD-specific antibodies can
be predetermined. The threshold refers to a level of the
RBD-specific antibodies above which the sample is scored as being
positive for RBD-specific antibodies. For example, the threshold
can be a level above which the sample is scored as having
sufficient neutralizing activity against the SARS-CoV-2. If the
level of the RBD-specific antibodies is below the threshold, it
could indicate insufficient protective immunity in the subject, and
hence likelihood of disease progression. In contrast, if the level
of the RBD-specific antibodies in the sample reaches or is above
the threshold, it could indicate protective immunity in the
subject, and hence less likely to suffer from disease
progression.
[0770] Any sample suspected of containing antibodies can be used.
In some embodiments, a suitable sample can be obtained from blood,
for example, a whole blood sample, a serum sample, or a plasma
sample. In some embodiments, said sample is obtained from a subject
suspected of having, inflicted with, or under treatment for
SARS-CoV-2 infection, or a disease, disorder or condition
associated with SARs-CoV-2 infection.
[0771] Polypeptides comprising the RBD of the spike protein of
SARS-CoV-2 can be used in the methods provided to herein to detect
presence or level of the RBD-specific antibodies in the subject. In
certain embodiments, the RBD of the spike protein of SARS-CoV-2
comprises an amino acid sequence comprising SEQ ID NO: 128. In
certain embodiments, the polypeptides can further comprise a tag.
Exemplary tag include, without limitation, 6.times. His tag or its
fusion such SEQ ID NO: 132 or SEQ ID NO: 133. The polypeptides
comprising RBD may be produced by recombinant methods (e.g., by
prokaryotic expression system or eukaryotic expression system), or
chemically synthesized (e.g. by solid phase synthesis, or solution
synthesis method). Solid phase synthesis method is described by
Merrifield in J.A.C.S. 85: 2149-2154 (1963) or the standard
solution synthesis method described in "Peptide Synthesis" by
Bodanszky, et al, second edition, John Wiley and Sons, 1976. The
polypeptides can be purified by methods known in the art. Various
methods of protein purification may be employed and such methods
are known in the art and described for example in Deutscher,
Methods in Enzymology, 182 (1990); Scopes, Protein Purification:
Principles and Practice, Springer-Verlag, New York (1982). The
purification step(s) selected will depend, for example, on the
nature of the production process used and the particular
polypeptide of the present application produced.
[0772] The presence or level of RBD-specific antibodies in a sample
can be determined based on the detection of the presence or level
of the complex of the RBD bound by the RBD-specific antibodies. Any
suitable methods can be used for such detection, for example, by
immunoassays such as immunohistochemistry immunofluorescence (IF),
immunoblotting (e.g., Western blotting), flow cytometry (e.g.,
FACS.TM.), Enzyme-linked Immunosorbant Assay (ELISA), enzyme
immunoassay (EIA), and radioimmunoassay (RIA), as described
above.
[0773] In certain embodiments, the polypeptide comprising RBD of
the spike protein of the SARS-CoV-2 may be immobilized on a solid
substrate. The immobilization can be via covalent linking or
non-covalent attachment (e.g. coating). The sample suspected of
containing the RBD-specific antibodies can be brought into contact
with the bound polypeptide. After a suitable period of incubation,
for a period of time sufficient to allow capture of the
RBD-specific antibodies via formation of antibody-antigen complex.
After washing away any unreacted materials, a detection antibody
specific to the captured antibody can be added, which can produce a
detectable signal to allow detection of the captured antibody. The
results may either be qualitative, by simple observation of the
visible signal, or may be quantitated by comparing with a control
sample containing known amounts of the detectable signal.
[0774] In another aspect, the present disclosure provides a method
of monitoring treatment response in a subject infected with
SARS-CoV-2 and received a treatment, the method comprising: (i)
contacting a sample from the subject with a peptide comprising an
amino acid sequence comprising SEQ ID NO: 128; (ii) detecting a
first level of an antibody in the sample wherein the antibody is
capable of specifically binding to RBD of the spike protein of the
SARS-CoV-2; and (iii) comparing the first level of the antibody
with a second level of the antibody detected in the subject prior
to the treatment; wherein the first level being higher than the
second level indicates that the subject is responsive to the
treatment.
[0775] In one embodiment, a sample is obtained from a subject or
patient prior to any treatment. In another embodiment, a test
sample is obtained during or after treatment such as anti-viral
treatment.
[0776] In one aspect, the present disclosure provides a kit for
detecting an antibody capable of specifically binding to
receptor-binding domain (RBD) of the spike protein of SARS-CoV-2,
comprising a polypeptide comprising an amino acid sequence
comprising SEQ ID NO: 128. In some embodiments, the polypeptide is
immobilized on a substrate. In some embodiments, the kit further
comprises a set of reagents for detecting complex of the antibody
bound to the polypeptide.
[0777] Kits
[0778] In certain embodiments, the present disclosure provides a
kit comprising one or more of the antibody or an antigen-binding
fragment thereof provided herein. In certain embodiments, the kit
disclosed herein is a therapeutic kit. In certain embodiments, the
kit disclosed herein is a diagnostic kit.
[0779] Such kits can further include, if desired, one or more of
various conventional kit components, such as, for example,
containers with one or more pharmaceutically acceptable carriers,
additional containers etc., as will be readily apparent to a person
skilled in the art. Instructions, either as inserts or a label,
indicating quantities of the components to be administered,
guidelines for administration, and/or guidelines for mixing the
components, can also be included in the kit.
[0780] In certain embodiments, where the antibody is labeled with
an enzyme, the kit will include substrates and cofactors required
by the enzyme (e.g., a substrate precursor which provides the
detectable chromophore or fluorophore). In addition, other
additives may be included such as stabilizers, buffers (e.g., a
block buffer or lysis buffer) and the like. The relative amounts of
the various reagents may be varied widely to provide for
concentrations in solution of the reagents which substantially
optimize the sensitivity of the assay. Particularly, the reagents
may be provided as dry powders, usually lyophilized, including
excipients which on dissolution will provide a reagent solution
having the appropriate concentration.
[0781] Also provided are diagnostic or detection reagents and kits
comprising one or more such reagents for use in a variety of
detection assays, including for example, immunoassays such as ELISA
(sandwich-type or competitive format). The kit's components may be
pre-attached to a solid support, or may be applied to the surface
of a solid support when the kit is used. In some embodiments, the
signal generating means may come pre-associated with an antibody of
the invention or may require combination with one or more
components, e.g., buffers, antibody-enzyme conjugates, enzyme
substrates, or the like, prior to use. Kits may also include
additional reagents, e.g., blocking reagents for reducing
nonspecific binding to the solid phase surface, washing reagents,
enzyme substrates, and the like. The solid phase surface may be in
the form of a tube, a bead, a microtiter plate, a microsphere, or
other materials suitable for immobilizing proteins, peptides, or
polypeptides. In particular aspects, an enzyme that catalyzes the
formation of a chemiluminescent or chromogenic product or the
reduction of a chemiluminescent or chromogenic substrate is a
component of the signal generating means. Such enzymes are well
known in the art. Kits may comprise any of the capture agents and
detection reagents described herein. Optionally the kit may also
comprise instructions for carrying out the methods of the
invention.
[0782] The detection kits disclosed herein may also be prepared
that comprise at least one of the antibodies or antigen-binding
fragments disclosed herein and instructions for using the
composition as a detection reagent. Containers for use in such kits
may typically comprise at least one vial, test tube, flask, bottle,
syringe or other suitable container, into which one or more of the
detection composition(s) may be placed, and preferably suitably
aliquoted. The kits disclosed herein will also typically include a
means for containing the vial(s) in close confinement for
commercial sale, such as, e.g., injection or blow-molded plastic
containers into which the desired vial(s) are retained. Where a
radiolabel, chromogenic, fluorigenic, or other type of detectable
label or detecting means is included within the kit, the labeling
agent may be provided either in the same container as the detection
composition itself, or may alternatively be placed in a second
distinct container means into which this second composition may be
placed and suitably aliquoted. Alternatively, the detection reagent
may be prepared in a single container means, and in most cases, the
kit will also typically include a means for containing the vial(s)
in close confinement for commercial sale and/or convenient
packaging and delivery.
[0783] A device or apparatus for carrying out the detection or
monitoring methods described herein is also provided. Such an
apparatus may include a chamber or tube into which sample can be
input, a fluid handling system optionally including valves or pumps
to direct flow of the sample through the device, optionally filters
to separate plasma or serum from blood, mixing chambers for the
addition of capture agents or detection reagents, and optionally a
detection device for detecting the amount of detectable label bound
to the capture agent immunocomplex. The flow of sample may be
passive (e.g., by capillary, hydrostatic, or other forces that do
not require further manipulation of the device once sample is
applied) or active (e.g., by application of force generated via
mechanical pumps, electroosmotic pumps, centrifugal force, or
increased air pressure), or by a combination of active and passive
forces.
[0784] The following examples are provided to better illustrate the
claimed invention and are not to be interpreted as limiting the
scope of the invention. All specific compositions, materials, and
methods described below, in whole or in part, fall within the scope
of the present invention. These specific compositions, materials,
and methods are not intended to limit the invention, but merely to
illustrate specific embodiments falling within the scope of the
invention. One skilled in the art may develop equivalent
compositions, materials, and methods without the exercise of
inventive capacity and without departing from the scope of the
invention. It will be understood that many variations can be made
in the procedures herein described while still remaining within the
bounds of the present invention. It is the intention of the
inventors that such variations are included within the scope of the
invention.
EXAMPLE 1
[0785] Materials and Methods
[0786] Patients and blood samples. A total of eight patients aged
10 to 66 years old infected with SARS-CoV-2 were enrolled (Table
5). A plasma sample from a healthy control was also included. Of
these eight patients, six (P#1 through P#4, P#8, and P#16) had
exposure history through personal visit and two had direct contact
with individuals from exposed area. Four subjects (P#1 through P#4)
were part of a family cluster (P#1 through P#5) infected and
subsequently transmitted infection to P#5 after returning to
Shenzhen. All patients were hospitalized at Shenzhen Third People's
Hospital, the designated city hospital for treatment of COVID-19
infected patients, three to nine days after symptom onset. All
patients presented with fever, fatigue, and dry cough and three
(P#1, P#2 and P#5) developed severe pneumonia. Four patients (P#1,
P#2, P#5, and P#22) were 60 years or older, of which three (P#1,
P#2, and P#22) had underlying disease such as hypertension.
SARS-CoV-2 infection status was verified by RT-PCR of
nasopharyngeal swab and throat swab specimens. No patient had
detectable influenza A, B, respiratory syncytial virus (RSV), or
adenovirus co-infections. Chest computed tomographic scans showed
varying degrees of bilateral lung patchy shadows or opacity. All
patients received interferon and ribavirin and/or
methylprednisolone treatments, recovered and were discharged except
for P#1, who succumbed to disease in hospital. Single (P#1, P#3,
P#5, P#8, P#16, and P#22) or sequential (P#2 and P#4) blood samples
were collected during hospitalization and follow-up visits and
separated into plasma and peripheral blood mononuclear cells
(PBMCs) by Ficoll-Hypaque gradient (GE Healthcare) centrifugation.
All plasma samples were heat-inactivated at 56.degree. C. for 1 h
before being stored at -80.degree. C. PBMCs were maintained in
freezing media and stored in liquid nitrogen until use.
[0787] Recombinant RBDs and trimeric Spike from SARS-CoV-2,
SARS-CoV, and MERS-CoV and receptor ACE2. Recombinant RBDs and
trimeric Spike for MERS-CoV, SARS-CoV, and SARS-CoV-2 and the
N-terminal peptidase domain of human ACE2 (residues Ser19-Asp615)
were expressed using the Bac-to-Bac baculovirus system (Invitrogen)
as previously described (Gui, M. et al. Cell Res 27, 119-129
(2017); Song, W. et al. PLoS Pathog 14, e1007236-e1007236 (2018);
Wang, N. et al. Cell Res 23, 986-993 (2013); Jiang, L. et al. Sci
Transl Med 6, 234ra259-234ra259 (2014); Zhang, S. et al. Cell Rep
24, 441-452 (2018)). Amino acid sequence for RBD of spike protein
for MERS-CoV is shown in SEQ ID NO: 126, and the polynucleotide
sequence is shown in SEQ ID NO: 127. Amino acid sequence for
extracellular domain of the spike protein for MERS-CoV is shown in
SEQ ID NO: 123. Amino acid sequence for RBD of spike protein for
SARS-CoV is shown in SEQ ID NO: 124, and the polynucleotide
sequence is shown in SEQ ID NO: 125. Amino acid sequence for
extracellular domain of the spike protein for SARS-CoV is shown in
SEQ ID NO: 122. Amino acid sequence for RBD of spike protein for
SARS-CoV-2 is shown in SEQ ID NO: 128, and the polynucleotide
sequence is shown in SEQ ID NO: 129. Amino acid sequence for
extracellular domain of the spike protein for SARS-CoV-2 is shown
in SEQ ID NO: 121. Extracellular domains of the spike protein were
fused to an artificial sequence to enable formation of a trimeric
Spike structure in vitro.
[0788] SARS-CoV-2 RBD (residues Arg319-Phe541) containing the gp67
secretion signal peptide (SEQ ID NO: 130) and a C-terminal
hexahistidine tag (SEQ ID NO: 132) or strap tag was inserted into
pFastBac-Dual vectors (Invitrogen) and transformed into DH10Bac
component cells. The bacmid was extracted and further transfected
into Sf9 cells using cationic lipid Cellfectin.RTM. II Reagents
(Invitrogen). The recombinant viruses were harvested from the
transfected supernatant and amplified to generate high-titer virus
stock. Viruses were then used to infect High Five cells for RBD and
trimeric Spike expression. Secreted RBD and trimeric Spike were
harvested from the supernatant and purified by gel filtration
chromatography as previously reported (Gui, M. et al. Cell Res 27,
119-129 (2017); Song, W. et al. PLoS Pathog 14, e1007236-e1007236
(2018); Wang, N. et al. Cell Res 23, 986-993 (2013); Jiang, L. et
al. Sci Transl Med 6, 234ra259-234ra259 (2014); Zhang, S. et al.
Cell Rep 24, 441-452 (2018)).
[0789] ELISA analysis of plasma and antibody binding to RBD,
trimeric Spike, and NP proteins. The recombinant RBDs and trimeric
Spike derived from SARS-CoV-2, SARS-CoV and MERS-CoV and the
SARS-CoV-2 NP protein (Sino Biological, Beijing) were diluted to
final concentrations of 0.5 .mu.g/ml or 2 .mu.g/ml, then coated
onto 96-well plates and incubated at 4.degree. C. overnight.
Samples were washed with PBS-T (PBS containing 0.05% Tween 20) and
blocked with blocking buffer (PBS containing 5% skim milk and 2%
BSA) at RT for 1 h. Either serially diluted plasma samples or
isolated mAbs were added the plates and incubated at 37.degree. C.
for 1 h. Wells were then incubated with secondary anti-human IgG
labeled with HRP (ZSGB-BIO, Beijing) and TMB substrate (Kinghawk,
Beijing) and optical density (OD) was measured by a
spectrophotometer at 450 nm and 630 nm. The serially diluted plasma
from healthy individuals or mAbs against SARS-CoV, MERS-CoV or
HIV-1 were used as controls.
[0790] Isolation of RBD-specific single B cells by FACS.
RBD-specific single B cells were sorted as previously described
(Kong, L. et al. Immunity 44, 939-950 (2016); Wu, X. et al. Science
329, 856-861 (2010)). In brief, PBMCs from infected and
convalescent individuals were collected and incubated with an
antibody and RBD cocktail for identification of RBD-specific B
cells. The cocktail consisted of CD19-PE-Cy7, CD3-Pacific Blue,
CD8-Pacific Blue, CD14-Pacific Blue, CD27-APC-H7, IgG-FITC (BD
Biosciences) and the recombinant RBD-Strep or RBD-His described
above. Three consecutive staining steps were conducted. The first
was a LIVE/DEAD Fixable Dead Cell Stain Kit (Invitrogen) in 50
.mu.l phosphate-buffered saline (PBS) applied at RT for 20 minutes
to exclude dead cells. The second utilized an antibody and RBD
cocktail for an additional 30 min at 4.degree. C. The third
staining at 4.degree. C. for 30 min involved either:
Streptavidin-APC (eBioscience) and/or Streptavidin-PE (BD
Biosciences) to target the Strep tag of RBD, or anti-his-APC and
anti-his-PE antibodies (Abcam) to target the His tag of RBD. The
stained cells were washed and resuspended in PBS before being
strained through a 70 .mu.m cell mesh (BD Biosciences).
RBD-specific single B cells were gated as
CD19+CD3-CD8-CD14-IgG+RBD+ and sorted into 96-well PCR plates
containing 20 .mu.l of lysis buffer (5 .mu.l of 5.times. first
strand buffer, 0.5 .mu.l of RNase out, 1.25 .mu.l of 0.1 M DTT
(Invitrogen) per well and 0.0625 .mu.l of Igepal (Sigma). Plates
were then snap-frozen on dry ice and stored at -80.degree. C. until
RT reaction.
[0791] Single B cell PCR, cloning and expression of monoclonal
antibodies (mAbs). The IgG heavy and light chain variable genes
were amplified by nested PCR and cloned into linear expression
cassettes or expression vectors to produce full IgG1 antibodies as
previously described (Liao, H.-X. et al. J Virol Methods, 2009;
Tiller, T.et al. J. Immunol Methods, 2008). Specifically, all
second round PCR primers containing tag sequences were used to
produce the linear Ig expression cassettes by overlapping PCR.
Separate primer pairs containing the specific restriction enzyme
cutting sites (heavy chain, 5'-AgeI/3'-SalI; kappa chain,
5'-AgeI/3'-BsiWI; and lambda chain, 5'-AgeI/3'-XhoI) were used to
amplify the cloned PCR products. The PCR products were purified and
cloned into the backbone of antibody expression vectors containing
the constant regions of human IgG1. The DNA sequence for the heavy
chain constant region of human IgG1 is set forth in SEQ ID NO: 118,
and the amino acid sequence for the heavy chain constant region of
human IgG1 is shown in SEQ ID NO: 115. Overlapping PCR products of
paired heavy and light chain expression cassettes were
co-transfected into 293T cells (ATCC) grown in 24-well plates.
Antigen-specific ELISA was used to detect the binding capacity of
transfected culture supernatants to SARS-CoV-2 RBD. Monoclonal
antibodies were produced by transient transfection of 293F cells
(Life Technologies) with equal amounts of paired heavy and light
chain plasmids.
[0792] Specifically, Table 4 shows the amino acid sequences and the
encoding DNA sequences for the heavy chain and light chain constant
regions of the monoclonal antibodies including P2A-1A8, P2A-1A9,
P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4, P2B-2G11, P2C-1A3, P2C- 1C8,
P2C-1C10, P2C-1D5, P2C-1F11, P2B-1G5, P2B-1A1, P2C-1D7, P2B-1A10,
P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2,
P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6,
P5A-1B8, P5A-1B9, P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9, P5A-2H3,
P5A-3A1, P5A-3A6, P5A-3B4, P5A-3C12, and P22A-1D1. Antibodies
P2A-1A8, P2A-1A9, P2B-2F6, P2B-2G4, P2B-2G11, P2C-1D5, P2B-1G5,
P2B-1A1, P2B-1D9, P2B-1E4, P5A-2G7, P5A-1D2, P5A-2E1, P5A-1D10,
P5A-2D11, P5A-2G9, P5A-2H3, P5A-3A6, and P5A-3B4 have lambda light
chains, and the amino acid sequence and encoding DNA sequence for
the lambda constant region are shown in SEQ ID NO: 116 and SEQ ID
NO: 119, respectively. Antibodies P2A-1A10, P2A-1B3, P2C-1A3,
P2C-1C8, P2C-1C10, P2C-1F11, P2C-1D7, P2B-1A10, P2B-1G1, P4A-2D9,
P5A-3C8, P5A-2F11, P5A-1C8, P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6,
P5A-1B8, P5A-1B9, P5A-1D1, P5A-3A1, P5A-3C12, and P22A-1D1have
kappa light chains, and the amino acid sequence and the encoding
DNA sequence for the kappa constant region are shown in SEQ ID NO:
117 and SEQ ID NO: 120, respectively.
[0793] Antibodies in the culture supernatant was purified by
affinity chromatography using Protein A beads columns (National
Engineering Research Center for Biotechnology, Beijing) according
to the manufacturer's protocol. Concentrations were determined by
BCA Protein Assay Kits (Thermo Scientific). SARS-CoV, MERS-CoV, and
HIV-1 mAbs were also included as controls. SARS-CoV antibodies
(S230 and m396) previously isolated by others (Zhu, Z. et al. Proc
Natl Acad Sci USA 104, 12123-12128 (2007)) were synthesized,
expressed in 293T cells and purified by protein A chromatography.
MERS-CoV antibodies (Mab-GD33) were derived from previously
reported (Niu, P. et al. J Infect Dis 218, 1249-1260 (2018)). HIV-1
antibody VRC01 was a broadly neutralizing antibody directly
isolated from a patient targeting the CD4 binding site of envelope
glycoprotein 40.
[0794] Antibody binding kinetics, epitope mapping, and competition
with receptor ACE2 measured by SPR. The binding kinetics and
affinity of mAbs to SARS-CoV-2 RBD were analyzed by SPR (Biacore
T200, GE Healthcare). Specifically, purified RBDs were covalently
immobilized to a CM5 sensor chip via amine groups in 10mM sodium
acetate buffer (pH 5.0) for a final RU around 250. SPR assays were
run at a flow rate of 30m1/min in HEPE buffer. The sensograms were
fit in a 1:1 binding model with BIA Evaluation software (GE
Healthcare). For epitope mapping, two different antibodies were
sequentially injected and monitored for binding activity to
determine whether the two mAbs recognized separate or
closely-situated epitopes. To determine competition with the human
ACE2 peptidase domain, SARS-CoV-2 RBD was immobilized to a CM5
sensor chip via amine group for a final RU around 250. Antibodies
(1 .mu.M) were injected onto the chip until binding steady-state
was reached. ACE2 (2 .mu.M), which was produced and purified as
above, was then injected for 60 seconds. Blocking efficacy was
determined by comparison of response units with and without prior
antibody incubation.
[0795] Analysis of plasma and antibody binding to cell surface
expressed trimeric Spike protein. HEK 293T cells were transfected
with expression plasmid encoding the full length spike of
SARS-CoV-2, SARS-CoV or MERS-CoV and incubated at 37.degree. C. for
36 h. The cells were removed from the plate using trypsin and
distributed into 96 well plates for the individual staining. Cells
were washed twice with 200 .mu.l staining buffer (PBS with 2%
heated-inactivated FBS) between each of the following. The cells
were stained at room temperature for 30 minutes in 100 .mu.l
staining buffer with 1:100 dilutions of plasma or 20 .mu.g/ml
monoclonal antibodies. The cells were then stained with PE labeled
anti-human IgG Fc secondary antibody (Biolegend) at a 1:20 dilution
in 50 .mu.l staining buffer at room temperature for 30 minutes.
Finally, the cells were re-suspended and analyzed with FACS Calibur
instrument (BD Biosciences, USA) and FlowJo 10 software (FlowJo,
USA). HEK 293T cells without transfection were also stained as
background control. S230 and m396 targeting the RBD of SARS-CoV
spike (Zhu, Z. et al. Proc Natl Acad Sci USA 104, 12123-12128
(2007)) and Mab-GD33 targeting the RBD of MERS-CoV spike (Niu, P.
et al. J Infect Dis 218, 1249-1260 (2018)) were used as positive
primary antibody controls, while VRC01 targeting HIV-1 env (Wu, X.
et al. Science 329, 856-861 (2010)) was used as an irrelevant
primary antibody control.
[0796] Neutralization activity of mAbs against pseudovirus and live
SARS-CoV-2. SARS-CoV-2, SARS-CoV and MERS-CoV pseudovirus were
generated by co-transfection of human immunodeficiency virus
backbones expressing firefly luciferase (pNL43R-E-luciferase) and
pcDNA3.1 (Invitrogen) expression vectors encoding the respective
full length S proteins into 293T cells (ATCC) (Wang, N. et al. Cell
Res 23, 986-993 (2013); Jiang, L. et al. Sci Transl Med 6,
234ra259-234ra259 (2014); Jia, W. et al. Emerg Microbes Infect 8,
760-772 (2019); Zhang, L. et al. J Med Virol 78, 1-8 (2006)). Viral
supernatants were collected 48 h later. Viral titers were measured
as luciferase activity in relative light units (Bright-Glo.TM.
Luciferase Assay Vector System, Promega Biosciences). Control
envelope glycoproteins derived from human immunodeficiency virus
(HIV)-1 and their corresponding pseudoviruses were produced in the
same manner. Control mAbs included VRC01 against HIV-1 40; S230 and
m396 against SARS-CoV (Zhu, Z. et al. Proc Natl Acad Sci USA 104,
12123-12128 (2007)); and Merb-GD33 against MERS-CoV 43.
Neutralization assays were performed by incubating pseudoviruses
with serial dilutions of purified mAbs at 37.degree. C. for 1 h.
Huh7 cells (ATCC) (approximately 1.5.times.10.sup.4 per well) were
added in duplicate to the virus-antibody mixture. Half-maximal
inhibitory concentrations (IC.sub.50) of the evaluated mAbs were
determined by luciferase activity 48 h after exposure to
virus-antibody mixture using GraphPad Prism 6 (GraphPad Software
Inc.), data were shown in Table 6 and Tables 7a, 7b and 7c.
[0797] Neutralization activity of mAbs against live SARS-CoV-2.
SARS-CoV-2 focus reduction neutralization test (FRNT) was performed
in a certified Biosafety Level 3 laboratory. Serial dilutions of
testing antibodies were conducted, mixed with 75 .mu.l of
SARS-CoV-2 (8.times.103 focus forming unit/ml, FFU/ml) in 96-well
microwell plates and incubated for 1 hour at 37.degree. C. Mixtures
were then transferred to 96-well plates seeded with Vero E6 cells
and allowed absorption for 1 hour at 37.degree. C. Inoculums were
then removed before adding the overlay media (100 .mu.l MEM
containing 1.6% Carboxymethylcellulose, CMC). The plates were then
incubated at 37.degree. C. for 24 hours. Cells were fixed with 4%
paraformaldehyde solution for 30 min, and overlays were removed.
Cells were permeabilized with 0.2% nonionic surfactant Triton.TM.
X-100 and incubated with cross-reactive rabbit anti-SARS-CoV-N IgG
(Sino Biological, Inc) for 1 hour at room temperature before adding
HRP-conjugated goat anti-rabbit IgG(H+L) antibody (Jackson
ImmunoResearch). Cells were further incubated at room temperature.
The reactions were developed with KPL TrueBlue Peroxidase
substrates (Seracare Life Sciences Inc). The numbers of SARS-CoV-2
foci were calculated using an EliSpot reader (Cellular Technology
Ltd).
[0798] Gene family usage and phylogenetic analysis of mAbs. The
program IMGT/V-QUEST (http://www.imgt.org/IMGT_vquest/vquest) was
used to analyze germline gene, germline divergence or degree of
somatic hypermutation (SHM), the framework region (FR) and the loop
length of the complementarity determining region 3 (CDR3) for each
antibody clone. The IgG heavy and light chain variable genes were
aligned using Clustal W in the BioEdit sequence analysis package
(https://bioedit.software.informer.com/7.2/). Phylogenetic analyses
were performed by the Maximum Likelihood method using MEGA X
(Molecular Evolutionary Genetics Analysis across computing
platforms). Several forms of the phylogenetic trees are presented
for clarity, data were shown in Table 9a, Table 9b, FIG. 4U and
FIG. 4V.
[0799] Antibody production. The production of antibodies was
conducted as previously described (Jiang, L. et al. Sci Transl Med
6, 234ra259-234ra259 (2014); Zhang, Q. et al. Sci Rep 6,
25856-25856 (2016)). The genes encoding the heavy and light chains
of isolated antibodies were separately cloned into expression
vectors containing IgG1 constant regions and the vectors were
transiently transfected into HEK293T or 293F cells using
polyethylenimine (PEI) (Sigma). After 72 h, the antibodies secreted
into the supernatant were collected and captured by protein A
SEPHAROSE.TM. (GE Healthcare). The bound antibodies were eluted and
further purified by gel-filtration chromatography using a
SUPERDEX.TM. 200 High Performance column (GE Healthcare). The
purified antibodies were either used in binding or neutralizing
assays.
[0800] Crystallization and data collection. The SARS-CoV-2 RBD was
mixed with the Fab fragment of P2B-2F6, P5A-1D2, P5A-3C8 or
P22A-1D1 respectively at a molar ratio of 1:1.2, incubated for 2 h
at 4.degree. C. and further purified by gel-filtration
chromatography. The purified complex concentrated to approximately
10 mg/mL in HBS buffer (10 mM HEPES, pH 7.2, 150 mM NaCl) was used
for crystallization. The screening trials were performed at
18.degree. C. using the sitting-drop vapor-diffusion method by
mixing 0.2 .mu.L of protein with 0.2 .mu.L of reservoir solution.
Crystals were successfully obtained in 0.2 M magnesium formate
dihydrate, 0.1M sodium acetate trihydrate, pH 4.0, 18% PEG5000mme.
The purified complexes of SARS-CoV-2 RBD and the Fab fragment of
P2C-1F11, P5A-1D2, P5A-3C8 or P22A-1D1 respectively were obtained
using a similar process. Crystals were successfully obtained in 0.2
M magnesium formate dihydrate, 0.1M sodium acetate trihydrate, pH
4.0, 18% PEG5000mme for P2C-1F11; in 0.2M Magnesium chloride
hexahydrate, 0.1M tris(hydroxymethyl)aminomethane buffer (Tris), pH
8.5, 3.4M 1,6-Hexanediol for P5A-1D2; in 0.2M Lithium sulfate
monohydrate, 0.1M HEPES, pH 7.5, 25% w/v PEG 3350 for P5A-3C8; and
in 0.1M potassium chloride, 0.1M NaHEPES, pH 7.0, 15% PEG 5000MME
for P22A-1D1, respectively. Crystals were harvested, soaked briefly
in mother liquid with 20% glycerol, and flash-frozen in liquid
nitrogen. Diffraction data was collected at 100 K and at a
wavelength of 0.97918 .ANG. on the BL17U beam line of the Shanghai
Synchrotron Research Facility (SSRF). Diffraction data was
auto-processed with aquarium pipeline and the data processing
statistics are listed in Table 10a and Table 10b and Table 10c.
(McCoy, A. J. et al. Journal of applied crystallography 40,
658-674, (2007)).
[0801] Structural determination and refinement. The structure was
determined by the molecular replacement method with PHASER in CCP4
suite (Cohen, S. X. et al. Acta crystallographica. Section D,
Biological crystallography 64, 49-60, (2008)). The search models
were the SARS-CoV-2 RBD structure (PDB ID: 6M0J) and the structures
of the variable domain of the heavy and light chains available in
the PDB with the highest sequence identities. Subsequent model
building and refinement were performed using COOT and PHENIX,
respectively (Emsley, P. & Cowtan, K. Acta crystallographica.
Section D, Biological crystallography 60, 2126-2132, (2004); Adams,
P. D. et al. Acta crystallographica. Section D, Biological
crystallography 58, 1948-1954, (2002)). Final Ramachandran
statistics: 90.02% favoured, 8.24% allowed and 1.74% outliers for
the final RBD-P2C-1F11 complex structure. Final Ramachandran
statistics: 95% favoured, 3.9% allowed and 0.81% outliers for the
final RBD-P22A-1D1 complex structure; Final Ramachandran
statistics: 94.23% favoured, 5.44% allowed and 0.32% outliers for
the final RBD-P5A-1D2 complex structure; Final Ramachandran
statistics: 97% favoured, 3.1% allowed and 0.33% outliers for the
final RBD-P5A-3C8 complex structure. The structural refinement
statistics are listed in Table 10a and Table 10b. All structural
figures were generated using PyMOL (Janson, G., Zhang, C., Prado,
M. G. & Paiardini, A. Bioinformatics (Oxford, England) 33,
444-446, (2017)).
[0802] Analysis of antibody binding to cell surface expressed
wild-type and mutant Spike protein. Single Alanine mutations were
conducted with QuickChange Site-directed mutagenesis Kit (Agilent
210518) followed the manufacturer's instructions. HEK 293T cells
were transfected with expression plasmid encoding either wild-type
or mutant full-length SARS-Cov-2 and incubated at 37.degree. C. for
36 h. The cells were removed from the plate using trypsin and
distributed into 96 well plates for the individual staining. Cells
were kept at 4.degree. C. or on ice in the following incubation or
wash steps. Cells were washed twice with 200 .mu.L ice-cold
staining buffer (PBS with 2% heated-inactivated FBS) between each
of the following. The cells were stained for 1 h in 100 .mu.L
staining buffer with 10 ug/mL ACE2 protein or 2 .mu.g/mL monoclonal
antibodies. The cells were then stained with one of the following
secondary antibodies: anti-his PE (Miltyni 130120787) for ACE2,
anti-human IgG Fc PE (Biolegend 410718) for nAbs, or anti-mouse IgG
Fc FTIC (ThermoFisher A10673) for S2 mAb (MP 08720401). Finally,
the cells were re-suspended and analyzed with FACS Calibur
instrument (BD Biosciences, USA) and FlowJo 10 software (FlowJo,
USA). HEK 293T cells without mock transfection were stained as
background control.
EXAMPLE 2
[0803] This example illustrates the identification of human plasma
and B cell that responses specific to SARS-CoV-2 RBD.
[0804] Cross-sectional and longitudinal blood samples from eight
SARS-CoV-2-infected and convalescent subjects were collected during
the early outbreak in Shenzhen (see Table 5). Samples were named by
patient number and either A, B, or C depending on collection
sequence. Six patients (P#1 through P#4, P#8, and P#16) had travel
history to exposed area and the remaining two (P#5 and P#22) had
direct contact with those from exposed area. P#1 through P#5 is a
family cluster with the first documented case of human-to-human
transmission of SARS-CoV-2 in Shenzhen. All subjects recovered and
were discharged from the hospital except for P#1 who succumbed to
disease despite intensive intervention. To analyze antibody
binding, serial plasma dilutions were applied to enzyme-linked
immunosorbent assay (ELISA) plates coated with either recombinant
RBD or trimeric Spike derived from SARS-CoV-2, SARS-CoV, and
MERS-CoV or recombinant NP from SARS-CoV-2. Binding activity was
visualized using anti-human IgG secondary antibodies at an optical
density (OD) of 450 nm. Varying degrees of binding were found
across individuals and among samples from the same individual.
Samples from P#1, P#2, P#5, and P#16 demonstrated higher binding to
both SARS-CoV-2 RBD and NP than the rest (FIG. 1A). Three
sequential plasma samples collected from P#2 over nine days during
early infection showed similar binding to SARS-CoV-2 RBD and NP and
remained relative stable over the course of the infection.
Surprisingly, virtually no cross-reactivity between SARS-CoV RBD
and MERS-CoV RBD was detected (FIG. 1A), despite strong recognition
by the positive control antibodies. However, strong
cross-reactivity was detected against trimeric Spikes from SARS-CoV
and MERS-CoV in both ELISA (FIG. 1B) and cell-surface staining
(FIG. 6A-FIG. 6C). All samples except P#4A demonstrated significant
levels of cross-binding to SARS-CoV trimeric Spike while only those
from P#1, P#2 and P#4B cross recognized MERS-CoV trimeric Spike
(FIG. 1B). None of the plasma samples were reactive to HIV-1
envelope trimer derived from strain BG505 (Sanders, R. W. et al. J
Virol 76, 8875-8889, (2002)). The same plasma samples were also
evaluated for neutralization of pseudoviruses bearing the Spike
proteins of either SARS-CoV-2, SARS-CoV, or MERS-CoV. Consistent
with the antibody binding results, varying degrees of neutralizing
activities against SARS-CoV-2 were found across individuals (FIG.
1C). However, cross-neutralizing against SARS-CoV and MERS-CoV is
rather minimal as all plasma samples tested, including healthy
control plasma, had negligible levels of neutralization (FIG. 1C).
No detectable neutralization was found for any plasma sample
against the pseudovirus control bearing the HIV-1 envelope MG04
(FIG. 1C). Taken together, these results suggest that RBDs from
SARS-CoV-2, SARS-CoV, and MERS-CoV are likely to be immunologically
distinct despite substantial sequence and structural similarities.
Thus, regions beyond RBDs likely contribute to the observed
cross-reactivity against SARS-CoV and MERS-CoV Spike protein.
[0805] Flow cytometry with a range of gating strategies was used to
study SARS-CoV-2-specific B cell responses and identity B cells
recognizing fluorescent-labeled RBD probes (FIG. 1D and FIG.
7A-FIG. 7K). As shown in FIG. 1E-FIG. 1F, the RBD-specific B cells
constitute about 0.005-0.065% among the total B cell population and
0.023-0.329% among the memory subpopulations. The number of
RBD-specific B cells are relatively higher in P#2, P#5, P#16, and
P#22 (FIG. 1E-FIG. 1F), which appeared to correlate well with
binding activity of corresponding plasma samples to SARS-CoV-2 RBD
and trimeric Spike protein (FIG. 1A and FIG. 1B). However, sample
P#1A demonstrated the lowest RBD-specific B cell response despite
high-level plasma binding. As P#1 was the only patient succumb to
disease, it is possible that this dichotomy of high plasma binding
activity and low levels of RBD-specific B cells is a surrogate
marker of rapid disease progression.
EXAMPLE 3
[0806] This example illustrates the cloning and analysis of single
B cell antibody against SARS-CoV-2 RBD.
[0807] The RBD-binding B cells identified in EXAMPLE 2 were
isolated into single cell suspension for cloning and evaluation of
the mAb response (FIG. 1D and FIG. 7A-FIG. 7K). Immunoglobulin
heavy and light chains were amplified by RT-PCR using nested
primers. The amplified products were cloned into linear expression
cassettes to produce full IgG1 antibodies as previously described
(Kong, L. et al. Immunity 44, 939-950 (2016); Liao, H.-X. et al. J
Virol Methods 158, 171-179). The number of B cell clones varied
from 10 to 10.sup.6 among the subjects (FIG. 8). Individual IgGs
were produced by transfection of linear expression cassettes and
tested for SARS-CoV-2 RBD reactivity by ELISA. On average,
fifty-eight percent of the antibody clones were reactive, although
great variability was found among different individuals (FIG. 8).
Out of 358 antibodies, 206 antibodies were found to specifically
bind to SARS-CoV-2 RBD, and by B cell cloning and sequencing, 165
distinct sequences were obtained (Table 9). These 206 antibodies
demonstrated significant differences in binding activity. For
example, a large number of antibodies from samples P#2B, P#2C,
P#4A, P4#B, P#5A, P#16A, and P#22A had OD 450 values well over 4.0,
while none of those from sample P#1A exceeded 4.0. There were too
few antibodies from P#3A and P#8A to make meaningful evaluations
(FIG. 8). Furthermore, samples from different study subjects also
demonstrated substantial differences in heavy chain variable gene
(VH) usage (FIG. 2). For instance, P#1 samples are dominated by
VH3-53, 3-13, and 1-69 which constituted approximately 21.4%,
14.3%, and 14.3% of the entire VH repertoire, respectively. Samples
from P#2 and P#5 are more diverse in distribution and frequency of
their VH usage. However, no single or group of VH families stood
out among study subjects, suggesting patients have immunologically
distinct responses to SARS-CoV-2 infection. This hypothesis is
supported by the phylogenetic analysis of all 206 VH sequences
superimposed with their corresponding binding activities as
presented in FIG. 2B. The high-binding clusters (80% of clusters
with OD 450>3) were widely distributed across multiple heavy
chain families. In fact, majority of the high-binding antibodies
were derived by clonal expansion of specific VH families in P#2,
P#4, and P#5. Similarly, the middle- (60-80% of clusters with OD
450>3) and low- (<60% cluster with OD 450>3) binding
clusters were also widely distributed and each consisted of
disproportionally represented VH gene families.
[0808] As P#2 showed a large number of RBD-binding antibodies and
was the only patient with three sequential blood samples, more
detailed characterization of P#2 antibodies were conducted. Among a
total of 69 antibodies from P#2, the majority (59%) were scattered
across various branches and the remaining (41%) were clonally
expanded into three major clusters (FIG. 3A). Antibodies from the
three time points (A, B, C) do not appear to group together but
rather interdigitate among themselves, suggesting they are highly
related during early infection. Three clones were significantly
enriched and each constituted between 12-14% of the entire tested
repertoire (FIG. 3A). Their heavy-chain variable regions belong to
the VH1-2*06, VH3-48*02, and VH3-9*01 families. The corresponding
light-chain kappa (lgk) belongs to 2-40*01/2D-40*01, 3-20*01, and
light-chain lambda (lgl) to 2-14*02 with the respective joining
segment kappa 4 (Jk4), Jk5 and joining segment lambda 1 (J11)
(Table 9). More importantly, these clonally expanded antibodies
were identified in all three samples indicating that they are
strongly selected for during infection. When comparing their
representation within each cluster, VH1-2*06 and VH3-9*01 appeared
to increase from approximately 33 to 45%, whereas VH3-48*02
decreased from 33 to 9% over the three time points, although the
number of clones was too small for statistical significance.
Interestingly, the somatic hypermutation (SHM) or germline
divergence for VH1-2*06 was 0% and this cluster persisted during
the study period. However, the SHM for VH3-48*02 reached as high as
9.6% and for VH3-9*01 reached 3.8% compared to the overall average
of 2.2%.+-.3.3% among the 69 VH sequences. Furthermore, the CDR3
length for VH1-2*06, VH3-48*02, and VH3-9*01 was 19aa, 16aa, and
23aa, respectively, compared with the overall average of 16.+-.4aa
among the 69 VH sequences. Close examination of the longest CDR3
from the VH3-9*01 cluster revealed richness in tyrosine, indicating
potential hydrogen bonding and hydrophobic interactions with the
surrounding residues. These results shed light on the clonal
expansion and broad diversity of RBD-specific antibodies during
early infection and their potential role in controlling SARS-CoV-2
infection.
[0809] Furthermore, we also conducted a genomic analysis and
compared the heavy chain variable gene (VH) and kappa or lambda
light chain variable (VK/VL) genes usage in the 13 mAbs (P22A-1D1,
P5A-1B9, P5A-2G7, P5A-2G9, P5A-1D1, P5A-1B8, P5A-1D2, P5A-3B4,
P5A-3C8, P5A-3C12, P2C-1F11, P2B-2F6 and P2B-1A10) with lowest
IC.sub.50 identified in the pesudovirus neutralizing analysis in
Example 6. Of these 13 mAbs, 7 were found to use IGHV3-53/3-66 and
paired predominantly with IGK1-9*01 (Table 9b). Four of the seven
were derived from P#5 (P5A-1D1, P5A-1B8, P5A-1D2, and P5A-3C8)
whereas two from P#2 (P2C-1F11 and P2B-1A10) and one from P#22
(P22A-1D1). Such high prevalence (53.8%) and from diverse
individuals among the top neutralizers indicated that IGHV3-53/3-66
represented one major and public antibody responses against
SARS-CoV-2 (FIG. 3B-FIG. 3E). This finding is consistent with
recent reports have also recognized disproportionally high
prevalence of IGHV3-53/3-66 among SARS-CoV-2 patients (Barnes et
al., 2020; Yuan et al., 2020). Furthermore, the CDR3 length of the
antibodies varied from 9 to 15, located in the shorter range among
the total 165 RBD-specific antibodies identified (FIG. 3F). Their
somatic hypermutation (SHM) were generally low and some reached 0%
for heavy chain (P22A-1D1) or light chain (P5A-1B8 and
P2C-1F11).
EXAMPLE 4
[0810] The present invention is further defined in the following
Examples. It should be understood that these Examples, while
indicating preferred embodiments of the invention, are given by way
of illustration only. From the above discussion and these Examples,
one skilled in the art can ascertain the essential characteristics
of this invention, and without departing from the spirit and scope
thereof, can make various changes and modifications of the
invention to adapt it to various uses and conditions.
[0811] This example illustrates the binding properties of the
antibodies against SARS-CoV-2 RBD.
[0812] Based on their representation and distribution on the
phylogenetic tree, 13 of the 69 P#2 antibodies sequences were
selected for further analysis (FIG. 3A, starred). Five P#1A
antibody clones were used as controls. Surface plasmon resonance
(SPR) with SARS-CoV-2 RBD showed that P#2 antibodies had
dissociation constants (Kd) ranging from 1.38 to 21.29 nM while
those from P#1 ranged from 8.48 to 260.50 nM or not detectable at
all (FIG. 4U and FIG. 9A-FIG. 9B). SHM did not appear to correlate
with Kd; some germline clones with 0% divergence in both VH and VL
genes (P2A-1A10, P2B-2G4, P2C-1A3, and P2C-1E1) had Kd values
ranging from 2.47 to 21.19 nM, which comparable to that (1.38 to
17.57 nM) of clones with higher levels of SHM (FIG. 4U). The Kd of
representative clones (P2A-1A8, P2A-1A10, and P2A-1B3) from the
three clonally expanded clusters span from 4.65 to 8.91 nM,
suggesting that their expansion may not be driven by affinity
maturation. Antibody P2B-1G5 was also tested for RBD binding, and
the Kd value was 0.1 nM (Table 7a). Next, each antibody for
competition with ACE2 for binding to the SARS-CoV-2 RBD were
measured (FIG. 4B, FIG. 4U, FIG. 4V and FIG. 10A-FIG. 10B).
Specifically, the RBD was covalently immobilized on a CMS sensor
chip and first saturated by antibody and then flowed through with
soluble ACE2. Competing capacity of each antibody was measured as
percent reduction in ACE2 binding with the RBD. As shown in (FIG.
4U, FIG. 10A, FIG. 10B), the evaluated antibodies demonstrated
various competing capacity with ACE2. The most powerful was
P2C-1F11. Two of the three representative antibodies from the
clonal expanded clusters (P2A-1A10 and P2A-1B3) had also strong
reduction. The third representative (P2A-1A8) only showed mild
reduction. Many antibodies had only limited competing power with
ACE2 despite impressive Kd values, suggesting binding affinity is
not predictive of ACE2 competing capacity. Antibody P2B-1G5 was
also tested for ACE2 competition, and showed 17.54% competition
with ACE2 (Table 7a). Control antibodies from P#1 demonstrated even
lower competing power with ACE2. Surprisingly, none of the
antibodies tested demonstrated cross-binding with SARS-CoV and
MERS-CoV RBD except P1A-1C7 (Kd=4.85 .mu.M), for which only limited
cross reactivity with SARS-CoV RBD was detected (FIG. 9A-FIG.
9F).
[0813] An additional set of 13 neutralizing antibodies were also
identified (see Example 5) These neutralizing antibodies
demonstrated high yet varying binding affinity to the SARS-CoV-2
RBD measured by surface plasmon resonance (SPR) (FIG. 9G and Table
9b). Most interestingly, of the top 13 neutralizing antibodies, 7
were found to use IGHV3-53/3-66 and paired predominantly with
IGK1-9*01 (Table 9b). Four of the seven were derived from P#5
(P5A-1D1, P5A-1B8, P5A-1D2, and P5A-3C8) whereas two from P#2
(P2C-1F11 and P2B-1A10) and one from P#22 (P22A-1D1) (FIG. 3D-3E).
Such high prevalence (53.8%) and from diverse individuals among the
top neutralizers indicated that IGHV3-53/3-66 represented one major
and public antibody responses against SARS-CoV-2. Furthermore,
their CDR3 length varied from 9 to 15, located in the shorter range
among the total 165 RBD-specific antibodies identified (FIG. 3F).
Their somatic hypermutation (SHM) were generally low and some
reached 0% for heavy chain (P22A-1D1) or light chain (P5A-1B8 and
P2C-1F11). Recent reports have also recognized disproportionally
high prevalence of IGHV3-53/3-66 among SARS-CoV-2 patients (Barnes
et al., 2020; Yuan et al., 2020).
[0814] These 13 mAbs demonstrated high yet varying binding affinity
to the SARS-CoV-2 RBD measured by surface plasmon resonance (SPR)
(FIG. 10C and Table 9b). All except P2B-1A10 displayed single digit
or less nanomolar binding affinity. Apart from P5A-3B4, these mAbs
shared strong competitive capacity with ACE2 in binding to
SARS-CoV-2 RBD, suggesting their potential mechanism of
neutralization (FIG. 10C and Table 9b).
EXAMPLE 5
[0815] This example illustrates the neutralizing properties of the
antibodies against pseudoviruses bearing the Spike protein of
SARS-CoV-2.
[0816] For a first set of antibodies P2A-1A8, P2A-1A9, P2A-1A10,
P2A-1B3, P2B-2F6, P2B-2G4, P2B-2G11, P2C-1A3, P2C-1C8, P2C-1C10,
P2C-1D5, and P2C-1F11, RBD binding and pseudoviruses neutralizing
activities were tested. Consistent with the competing capacity
findings, neutralizing activity varied considerably with IC.sub.50
values ranging from 0.03 to >50 .mu.g/ml (FIG. 4C-FIG. 4M).
Within this first set of antibodies, P2C-1F11, P2B-2F6 and P2C-1A3
were the most potent with IC.sub.50 0.03, 0.05, and 0.63 .mu.g/ml,
respectively. Overall, ACE2 competing capacity correlated well with
the neutralizing activities, although this correlation was not
exact in some instances. Notably, no cross-neutralization was found
either against pseudoviruses bearing the full length Spike of
SARS-CoV or MERS-CoV or with cell-surface staining of trimeric
SARS-CoV and MERS-CoV Spike (FIG. 11A-FIG. 11B).
[0817] Antibody P2B-1G5 was also tested for pseudoviruses
neutralizing activities, with an IC.sub.50 value of 0.11 .mu.g/ml.
The results are shown in Table 7a.
[0818] Pseudoviruses neutralizing activities were further tested
using a second set of antibodies P5A-2G7, P5A-3C8, P5A-1D2,
P2B-1G1, P5A-1C8, P5A-2F11, P5A-2E1, P2B-1A1, P2C-1D7, P2B-1A10,
P2B-1D9, P2B-1E4, and P4A-2D9. Results showed that most of these
antibodies were potent, and IC.sub.50 was found below 1 .mu.g/ml
for antibodies P2B-1G5, P5A-2G7, P5A-3C8, P5A-1D2, P5A-1C8,
P5A-2F11, P2B-1A1, P2C-1D7, and P2B-1A10 (Table 7b).
[0819] Pseudoviruses neutralizing activities were also tested using
a third set of antibodies P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6,
P5A-1B8, P5A-1B9, P5A-1D1, P5A-1D10, P5A- 2D11, P5A-2G9, P5A-2H3,
P5A-3A1, P5A-3A6, P5A-3B4, P5A-3C12, and P22A-1D1. Results showed
that most of these antibodies were potent, and IC.sub.50 was found
below 1 .mu.g/ml for antibodies P4A-1H6, P5A-1B6, P5A-1B8, P5A-1B9,
P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9, P5A-2H3, P5A-3A1, P5A-3A6,
P5A-3B4, P5A-3C12, and P22A-1D1 (Table 7c and FIG. 4V). Among them,
P5A-1B9, P22A-1D1, P5A-1D1, P5A-1B8, P5A-2G9, P5A-3B4 and P5A-3C12
were the most potent with IC.sub.50 lower than 0.1 .mu.g/ml
(0.0014, 0.0038, 0.0096, 0.0115, 0.0158, 0.0993 and 0.0996
.mu.g/ml, respectively).
[0820] By summarizing the initial screening result by using
pseudovirus, we identified 13 mAbs (P22A-1D1, P5A-1B9, P5A-2G7,
P5A-2G9, P5A-1D1, P5A-1B8, P5A-1D2, P5A-3B4, P5A-3C8, P5A-3C12,
P2C-1F11, P2B-2F6 and P2B-1A10) with IC.sub.50 ranging from 0.0014
.mu.g/mL to 0.0996 .mu.g/mL (FIG. 4N through FIG. 4R). The
IC.sub.50 of remaining antibodies, however, spans between 0.1
.mu.g/mL and 50 .mu.g/mL or higher (Table 7d).
[0821] We selected the top seven potent neutralizing antibodies
against pseudovirus in the first set of antibodies to analyze their
inhibitory activities against live SARS-CoV-2 using focus reduction
neutralization test (FRNT) (FIG. 4S) and FIG. 12A-FIG. 12D).
Consistent with their respective pseudovirus assay findings,
P2C-1F11, P2B-2F6 and P2C-1A3 demonstrated the most potent
neutralization activity with IC.sub.50 0.03 0.41, and 0.28
.mu.g/ml, respectively (FIG. 4U). The remaining antibodies
demonstrated moderate neutralizing activities with IC.sub.50
ranging from 1.64 to 35.87 .mu.g/ml (FIG. 4U). The further
identified top 13 neutralizing antibodies also demonstrated strong
inhibitory activity against live SARS-CoV-2 based on focus
reduction neutralization tests (FRNT) (FIG. 4T). For instance, the
IC.sub.50 for the best antibody P5A-1B9 reached as low as 0.0043
.mu.g/mL and the IC.sub.80 0.0441 .mu.g/mL, at least 10-fold more
potent than those tested in the first set (FIG. 4V).
[0822] To determine whether these antibodies compete for similar
epitopes on the SARS-CoV-2 RBD, a total of six antibodies with
relative strong ACE2 competitive capacities and neutralization
potency and analyzed in a pairwise competition fashion using SPR.
As shown in Table 8 and FIG. 13, variable degrees of competition
were found among the pairs of antibodies. P2C-1A3, for instance,
was competitive against all antibodies tested with strong reduction
capacity (FIG. 13). P2C-1F11, on the other hand, was less
competitive with other antibodies and in particular, only minimally
competitive with P2C-1C10. P2B-2F6, another potent neutralizing
antibody, was broadly competitive with all antibodies tested. These
results indicate that the antibodies analyzed recognized both
overlapping and distinct epitopes. Different mAbs may therefore
exert their neutralizing activity through different mechanisms.
[0823] Antibody P2B-1G5 was also analyzed in a pairwise competition
fashion with P2C-1F11 using SPR. Results was shown in Table 7a,
which suggested that P2B-1G5 is only minimally competitive with
P2C-1F11.
EXAMPLE 6
[0824] This example illustrates the structural basis for antibody
neutralization.
[0825] The crystal structure of the top three most potent
neutralizing antibodies in the first set of antibodies (P2C-1F11,
P2B-2F6 and P2C-1A3) was determined. Of which, P2B-2F6 Fab and
P2C-1F11 Fab bound to the SARS-CoV-2 RBD were able to form crystals
and the structures of which were resolved at a resolution of 2.85
angstrom (FIG. 5A and FIG. 5F).
[0826] Antibody 2F6 mainly uses the heavy chain for interactions
with the RBD, and the paratope region consists of 14 residues from
the heavy chain (Y27, S28, S30, S31 and Y33 of HCDR1; H54 of HCDR2;
G102, I103, V105, V106 and P107 of HCDR3) and 3 residues from the
light chain (G31, Y32 and N33 of LCDR1) (FIG. 5E). The buried
surface area on the RBD is 534 A.sup.2 and the recognized epitope
residues are all from the receptor-binding motif (RBM) of the RBD,
including residues K444, G446, G447, N448, Y449, N450, L452, V483,
E484, G485, F490 and S494 (FIG. 5E). SARS-CoV-2 recognition by 2F6
is largely driven by hydrophobic interactions around RBD residues
Y449, L452 and F490 (FIG. 5B). Structural superimposition of the
RBD-2F6 and RBD-ACE2 crystal structures indicated that the binding
of 2F6 would clash with ACE2 (FIG. 5C). The clash would happen
between the P2B-2F6 light chain (residues R56, S58, G59, R63, S78,
G79) and the ACE2 (residues D67, K68, A71, K74, E110, K114). The
overlapping residues recognized by 2F6 and ACE2 only include G446
and Y449, largely due to their difference in angles when they
approach RBD. However, the high affinity binding of 2F6 to the RBD
(5.14 nM), which is comparable to the binding affinity between RBD
and ACE2 (4.70 nM), is expected to preclude the receptor ACE2
engagement, further supported by the high ACE2 competition
efficiency of 2F6 in the SPR analysis (98.80% in FIG. 4O), second
column). We also superimposed the RBD-2F6 crystal structure onto
the cryo-EM structure of the SARS-COV-2 spike trimer, in which the
RBD has two different "up" and "down" conformations. Unlike the
ACE2 that only binds the "up" RBD, the 2F6 Fab is able bind to the
RBD in both "up" and "down" conformations without clashing with two
other monomers in the spike trimer (FIG. 5D). Therefore, we suggest
that structural basis for 2F6 neutralization relies on directly
competition with receptor ACE2 on spike binding.
[0827] Antibody 1F11 mainly uses the heavy chain for interactions
with the RBD, and the paratope region consists of 17 residues from
the heavy chain (G26, I27, T28, S31, N32 and Y33 of HCDR1; Y52,
S53, G54, and S56 of HCDR2; Y58 of HFR3; R97, L99, V100, V101, Y102
and D105 of HCDR3) and 4 residues from the light chain (12 from the
LFR1; S28, S30 and Y33 of LCDR1) (FIG. 5G). The buried surface area
on the RBD is shown in FIG. 5G and the recognized 23 epitope
residues are located in the RBM (Y453, L455, F456, R457, K458,
S459, N460, Y473, A475, G476, 5477, F486, N487, Y489, Q493, G502
and Y505) and the core (R403, T415, G416, K417, D420 and Y421) of
the SARS-CoV-2 RBD (FIG. 5G). A network of hydrogen-bonding
interactions (18 between heavy chain and RBD and 2 between light
chain and RBD) dominates in the recognition of SARS-CoV-2 by
1F11.
[0828] The crystal structures of P22A-1D1, P5A-3C8, and P5A-1D2
complexed with SARS-CoV-2 RBD (FIG. 5H and Table 10c) were also
determined at a resolution of 2.40 .ANG., 2.36 .ANG., and 2.60
.ANG. respectively. Antibody P2C-1F11 (2.96 .ANG.) was used it for
head to head comparison. As shown in FIGS. 5H, 5I, 5J, and 5K,
these four antibodies (P22A-1D1, P5A-3C8, P5A-1D2 and P2C-1F11)
bound to the RBD with a nearly identical angle of approach. The
estimated clash volume with ACE2 was about .about.20,000
.ANG..sup.3 (FIG. 5H), consistent with biochemical data showing
strong capacities to compete with ACE2 in binding to SARS-CoV-2 RBD
(Table 9b). The heavy chains of antibodies P22A-1D1, P5A-3C8,
P5A-1D2 and P2C-1F11 share similar buried surfaces on the RBD. The
estimated areas are 726 .ANG..sup.2 for P22A-1D1, 668 .ANG..sup.2
for P5A-3C8, 823 .ANG..sup.2 for P5A-1D2 and 725 .ANG..sup.2 for
P2C-1F11 (FIG. 5I). In contrast, the buried surface areas of the
light chain are rather different. P22A-1D1 (413 .ANG..sup.2) and
P5A-3C8 (480 .ANG..sup.2) are significantly larger than P5A-1D2
(152 .ANG..sup.2) and P2C-1F11 (230 .ANG..sup.2) (FIG. 5I). The
larger buried areas are translated into more epitope residues. For
instance, P22A-1D1 and P5A-3C8 have 28 and 31 epitope residues on
the RBD whereas P5A-1D2 and P2C-1F11 have 22 and 23, respectively
(FIG. 5S). Furthermore, the epitopes of these antibodies
significantly overlap with the ACE2 binding residues on RBD. Out of
17 ACE2-binding residues on RBD, P22A-1D1 shared by 15, P5A-3C8 by
16, P5A-1D2 by 10, and P2C-1F11 by 11 (FIG. 5L). The similar angles
of approach to and the large overlap in binding residues on the RBD
suggest that these four public antibodies resemble ACE2 in binding
to SARS-CoV-2. The coordinates and structure factor files for the
P5A-1D2, P5A-3C8 and P22A-1D1/SARS-CoV-2 RBD complexes have been
deposited in the Protein Data Bank (PDB) under accession numbers
7CHO, 7CHP, 7CHS, respectively.
[0829] As described in Example 3, the four antibodies P22A-1D1,
P5A-3C8, P5A-1D2 and P2C-1F11 were all found to use IGHV3-53 or
IGHV3-66 (Table 9b). The IGHV3-53 and IGHV3-66 share the identical
germline amino acid sequence except one residue. It is therefore
expected that the four antibodies shared their binding features to
RBD primarily through residues in the heavy chain. As shown in FIG.
5M, all three HCDRs are involved in the binding of these four
antibodies to the RBD. Heavy chain sequence alignments showed that
the HCDR1 and HCDR2 are highly conserved, whereas the HCDR3 are
rather different (FIG. 5P-FIG. 5S). Of note, P5A-1D2 has a longer
HCDR3 (15 residues) than the rest three antibodies (11
residues).
[0830] Antibody P22A-1D1 mainly uses the heavy chain for
interactions with the RBD, and the paratope region consists of 17
residues from the heavy chain (G26, F27, T28, S31, N32, Y33, H52,
S53, G54, S56, Y58, R97, R99, D100, Y101, Y102 and D105) and 10
residues from the light chain (Q27, G28, 129, S30, Y32, S67, H90,
L91, N92 and Y94) (FIG. 5T). The buried surface area on the RBD is
shown in FIG. 5T and the recognized 18 epitope residues are located
in the SARS-CoV-2 RBD (T415, G416, K417, D420, Y421, L455, F456,
R457, K458, N460, Y473, A475, G476, 5477, F486, N487, Y489 and
Q493) (FIG. 5T).
[0831] Antibody P5A-1D2 mainly uses the heavy chain for
interactions with the RBD, and the paratope region consists of 20
residues from the heavy chain (G26, F27, 128, S31, N32, Y33, Y52,
S53, G54, S56, Y58, R87, R97, L99, Q100, V101, G102, A103, T104 and
D106) and 3 residues from the light chain (A31, Y33, S95) (FIG.
5T). The buried surface area on the RBD is shown in FIG. 5T and the
recognized 20 epitope residues are located in the SARS-CoV-2 RBD
(T415, G416, K417, D420, Y421, Y453, L455, F456, R457, K458, N460,
Y473, Q474, A475, G476, S477, N487, Y489, Q493 and Y505) (FIG.
5T).
[0832] Antibody P5A-3C8 mainly uses the heavy chain for
interactions with the RBD, and the paratope region consists of 16
residues from the heavy chain (G26, F27, T28, S31, N32, Y33, Y52,
S53, G54, S56, Y58, R97, L99, Q100, E101 and H102) and 12 residues
from the light chain (G28, 129, S30, S31, S67, G68, H90, L91, N92,
S93 and Y94) (FIG. 5T). The buried surface area on the RBD is shown
in FIG. 5T and the recognized 19 epitope residues are located in
the SARS-CoV-2 RBD (T415, G416, K417, D420, Y421, Y453, L455, F456,
R457, K458, N460, Y473, A475, G476, S477, F486, N487, Y489 and
Q493) (FIG. 5T).
[0833] Antibody P2C-1F11 mainly uses the heavy chain for
interactions with the RBD, and the paratope region consists of 16
residues from the heavy chain (G26, 127, T28, S31, N32, Y33, Y52,
S53, G54, S56, R97, L99, V100, V101, Y102 and D105) and 3 residues
from the light chain (S28, S30 and Y33) (FIG. 5T). The buried
surface area on the RBD is shown in FIG. 5T and the recognized 19
epitope residues are located in the SARS-CoV-2 RBD (T415, G416,
K417, D420, Y421, L455, F456, R457, K458, N460, Y473, Q474, A475,
G476, S477, F486, N487, Y489 and Q493) (FIG. 5T).
[0834] In the shared HCDR1-RBD interface, the conserved HCDR1
residues G26, F27, T28/I28, S31, N32 and Y33 interact with RBD
residues L455, K458, Y473, A475, G476, S477 and N487. In the shared
HCDR2-RBD interface, interactions are largely mediated through
HCDR2 residues Y52, S53, G54, S56 and Y58 and RBD residues T415,
G416, K417, D420, Y421, K458 and N460. In particular, one unique
feature shared by the four antibodies is the participation of three
conserved tyrosines (Y33, Y52 and Y58) in forming a network of
hydrophobic and hydrophilic interactions with the RBD (FIG. 5N).
For example, the Y33 forms extensive hydrophobic interactions with
RBD K417, Y421, L455 and F456 (FIG. 5N). Its side chain --OH also
forms a conserved hydrogen bond with the main chain oxygen atom of
RBD L455 (FIG. 5N). Another unique and shared feature is the
interactions mediated by the -SGGS- segment in the HCDR2. Apart
from the close contacts through Van der Waals forces, specific
hydrogen-bonding interactions also occur between the beginning S53
and ending S56 with RBD Y421 and D420, respectively (FIG. 5O). In
addition, RBD Y421 also forms a conserved hydrogen bond with main
chain N atom of the G44 (FIG. 5O).
[0835] Despite of common and shared features, the four antibodies
also demonstrated some minor differences due to their sequence and
structure variations. P22A-1D1, P5A-3C8, and P2C-1F11 have the same
11-residue long HCDR3, but actual sequence varies. For example, the
-RDYYG- in P22A-1D1 is replaced by -LQEHG- in P5A-3C8 and by
-LVVYG- in P2C-1F11 (Table 9b). Therefore, although interacting
with the same RBD residues such as F456, N487, Y489 and Q493, the
specific residues in the HCDR3 in mediating such interactions are
different. Compared to the other three, P5A-1D2 has a relatively
longer HCDR3 with 15 residues (FIG. 5K and Table 9b), providing
more residues to interact with RBD. For instance, the T104 at the
tip of the P5A-1D2 HCDR3 has interactions with RBD Y505, which is
absent in other three HCDR3-RBD interfaces (FIG. 5K). RBD Y505 is
instead recognized by the light chain of P22A-1D1, P5A-3C8 and
P2C-1F11, and appears to serves as an anchor residue for light
chain binding (FIG. 5P-FIG. 5S). However, recognition by the long
HCDR3 of P5A-1D2 resulted in significant change in the side chain
conformation of Y505, precluding Y505 serving as an anchor for
P5A-1D2 light chain binding (FIG. 5P-FIG. 5S).
[0836] To further dissect the impact of epitope residues on the
binding of public antibodies, we conducted single-site alanine
scanning mutagenesis for the 15 epitope residues shared among the
public antibodies. All mutant spikes were successfully expressed on
the surface of HEK 293T cells, as measured by the median
fluorescence intensity (MFI) of the control S2 antibody through
flow cytometry. However, of the 15 mutant residues, 12 resulted in
more than 80% reduction in the binding of the four public
antibodies although some antibodies are more sensitive than others
(FIG. 6C, highlighted in grey boxes). For example, Y421A and F456A
have broad impact on all four public antibodies, whereas T415A,
Y473A, and N487A on three of the four. On the other hand, K417A,
D420A, L455A, R457A, N460A, and Y489A only reduced binding for two
of the four antibodies. In particular, Y505A appears to have more
profound impact on P5A-1D2 than the rest three antibodies. This is
perhaps due to the importance of Y505 in the heavy chain
recognition through T104 at the tip of the HCDR3 as illustrated
above. Taken together, these results indicate that some minor
differences do exist among the four public antibodies despite of
their overall similarity, which may account for their minor
differences in binding and neutralizing activities. Lastly, 9 out
of the 15 mutant residues also resulted in significant reduction of
ACE2 binding to the surface expressed spike glycoprotein. These
residues are highlighted in orange boxes including T415A, Y421A,
L455A, F456A, R457A, Y473A, N487A, Y489A, and Y505A. The shared
impact of these residues on ACE2 and the public antibodies support
the abovementioned structural analysis (FIG. 5H-FIG. 5L).
EXAMPLE 7
[0837] Half-Life and Human PK Results in Healthy Adult
Subjects.
[0838] The half-life of the monoclonal antibody (mAb) in a typical
70 kg subject was determined:
[0839] mAb T.sub.1/2=23.2 days (mAb with wild type human constant
domain Fc)
[0840] mAb-YTE T.sub.1/2=89.5 days (mAb with the YTE constant
domain Fc)
[0841] The mAb1 and mAb2 antibodies were constructed and produced
to contain the antigen-binding domain P2C-1F11 and P2B-1G5,
respectively. Both antibodies contain the modified human IgG
constant domain comprising a substitution with tyrosine at amino
acid residue 252, a substitution with threonine at amino acid
residue 254, and a substitution with glutamic acid at amino acid
residue 256, numbered according to the EU index as in Kabat.
[0842] Healthy human adult subjects were administered with the
antibody at specific dosages after obtaining consent according to
protocols approved by the institutional review board (IRB) (IRB
approval number (025)-02 and (026)-02, Ethical Committee of the
Beijing Ditan Hospital, Capital Medical University). Blood samples
were taken at indicated time points. Quantitation of mAb1 and mAb2
serum concentrations was conducted using a validated sandwich ELISA
method using a commercially available His-tag SARS-CoV-2 Spike
protein receptor binding domain as a capture reagent, and a
commercially available mouse Anti-Human IgG Fc antibody labelled
with horseradish peroxidase as detection reagent.
[0843] Three dose levels were performed: cohort 1 (750 mg), cohort
2 (1500 mg) and cohort 3 (3000 mg).
[0844] Using the established human population PK model, mAb1 and
mAb2 PK profiles at different dose levels were measured and
predicted using Monte Carlo simulation. The predicted medians are
shown in FIG. 15A-FIG. 15B as solid lines for each cohort. Dashed
lines and the dots represent measured concentrations in the
subjects. The shaded areas represent the 5th-95th percentiles.
EXAMPLE 8
[0845] Neutralization Effect.
[0846] The neutralizing activity of the mAb1 and mAb2 alone or in
combination against live virus SARS-CoV-2 were measured using focus
reduction neutralization test (FRNT) method. The two antibodies
exhibited potent antiviral activity and the combination of two
antibodies demonstrated a moderate additive effect in neutralizing
live SARS-CoV-2 virus (FIG. 16).
EXAMPLE 9
[0847] In Vivo Pharmacokinetics Assay.
[0848] The mAb1 and mAb2 single-dose PK was characterized after a
60-minutes IV infusion administration of the mAb1 and mAb2,
respectively, to male and female naive cynomolgus monkeys at 10
mg/kg (n=3 animals per sex group). The animal study was conducted
under the approved protocols.
[0849] Blood samples were collected at pre-dose, 5 min, 30 minutes,
1 hour, 2 hours, 4 hours, 8 hours, 24 hours, 48 hours, 72 hours, 96
hours, 120 hours, 168 hours, 240 hours, 336 hours, 504 hours, 672
hours, 840 hours, 1008 hours, 1176 hours and 1344 hours post-end of
infusion. Anti-drug antibody (ADA) samples were collected at
pre-dose, 336 hours, 504 hours, 672 hours, 840 hours, 1008 hours,
1176 hours, and 1344 hours post-end of infusion. Concentrations of
mAb1 or mAb2 in serum samples were determined by an ELISA method.
The level of ADA responses in collected samples were determined
using a validated ECL method. Pharmacokinetic non-compartmental
analyses (NCA) were based on the time of IV infusion initiation
time,
[0850] Pharmacokinetics (PK) data for the mAb1 and mAb2 in
Cynomolgus Monkey are shown in FIG. 17A (mAb1) and FIG. 17B (mAb2).
mAb1 showed no marked sex differences at the dose level. Serum mAb1
and mAb2 concentrations declined in a biphasic manner after a
single 10 mg/kg IV dose, with a concentration-time profile that
indicated linear elimination.
[0851] All of the compositions and methods disclosed and claimed
herein can be made and executed without undue experimentation in
light of the present disclosure. While the compositions and methods
of this invention have been described in terms of preferred
embodiments, it will be apparent to those of skill in the art that
variations may be applied to the methods and in the steps or in the
sequence of steps of the method described herein without departing
from the concept, spirit and scope of the invention. More
specifically, it will be apparent that certain agents which are
both chemically and physiologically related may be substituted for
the agents described herein while the same or similar results would
be achieved. All such similar substitutes and modifications
apparent to those skilled in the art are deemed to be within the
spirit, scope and concept of the invention as defined by the
appended claims.
Numbered Embodiments of the Disclosure
[0852] Other subject matter contemplated by the present disclosure
is set out in the following numbered embodiments: [0853] 1. An
isolated or recombinant antibody or an antigen-binding fragment
thereof, which is capable of specifically binding to SARS-CoV-2,
and exhibiting at least 50% less binding or non-detectable binding
to SARS-CoV or MERS-CoV. [0854] 2. An isolated or recombinant
antibody or an antigen-binding fragment thereof, having one or more
features selected from the group consisting of:
[0855] a) capable of specifically binding to spike protein of
SARS-CoV-2 and exhibiting at least 50% less binding to spike
protein of SARS-CoV or spike protein of MERS-CoV;
[0856] b) capable of specifically binding to receptor-binding
domain (RBD) of the spike protein of SARS-CoV-2 comprising the
amino acid sequence of SEQ ID NO: 128;
[0857] c) exhibiting binding to RBD of spike protein of SARS-CoV
comprising the amino acid sequence of SEQ ID NO: 124 at a level
that is non-detectable or that is no more than 50% of the binding
to the RBD of spike protein of SARS-CoV-2;
[0858] d) exhibiting binding to RBD of spike protein of MERS-CoV
comprising the amino acid sequence of SEQ ID NO: 126 at a level
that is non-detectable or that is no more than 50% of the binding
to RBD of the spike protein of SARS-CoV-2;
[0859] e) capable of binding to the RBD of spike protein of
SARS-CoV-2 at a K.sub.d value of no more than 1.times.10.sup.-7M as
measured by Surface Plasmon Resonance (SPR);
[0860] f) exhibiting binding to the RBD of spike protein of
SARS-CoV or the RBD of spike protein of MERS-CoV at a K.sub.d value
of at least 1.times.10.sup.-6M as measured by SPR;
[0861] g) capable of exhibiting at least 30% competition at with 2
.mu.M angiotensin converting enzyme 2 (ACE2) receptor, for binding
to the RBD of spike protein of SARS-CoV-2 immobilized at a
resonance units (RU) of 250, as measured by SPR; and
[0862] h) capable of binding to the RBD of spike protein of
SARS-CoV-2 at a neutralizing activity at an IC.sub.50 value of no
more than 100 .mu.g/ml, as measured by pseudovirus neutralization
assay. [0863] 3. An isolated or recombinant antibody or an
antigen-binding fragment thereof capable of specifically binding to
RBD of spike protein of SARS-CoV-2, comprising: [0864] a) 1, 2, or
3 heavy chain CDR sequences selected from SEQ ID NO: 1, SEQ ID NO:
2, and SEQ ID NO: 3; [0865] b) 1, 2, or 3 heavy chain CDR sequences
selected from SEQ ID NO: 11, SEQ ID NO: 12, and SEQ ID NO: 13;
[0866] c) 1, 2, or 3 heavy chain CDR sequences selected from SEQ ID
NO: 21, SEQ ID NO: 22, and SEQ ID NO: 23; [0867] d) 1, 2, or 3
heavy chain CDR sequences selected from SEQ ID NO: 31, SEQ ID NO:
32, and SEQ ID NO: 33; [0868] e) 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 41, SEQ ID NO: 42, and SEQ ID
NO: 43; [0869] f) 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 51, SEQ ID NO: 52, and SEQ ID NO: 53; [0870] g) 1,
2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 65, SEQ
ID NO: 66, and SEQ ID NO: 67; [0871] h) 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 75, SEQ ID NO: 76, and SEQ ID
NO: 77; [0872] i) 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 85, SEQ ID NO: 86, and SEQ ID NO: 87; [0873] j) 1,
2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 95, SEQ
ID NO: 96, and SEQ ID NO: 97; [0874] k) 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 105, SEQ ID NO: 106, and SEQ ID
NO: 107; [0875] l) 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 136, SEQ ID NO: 137, and SEQ ID NO: 138; [0876] m)
1, 2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 146,
SEQ ID NO: 147, and SEQ ID NO: 148; [0877] n) 1, 2, or 3 heavy
chain CDR sequences selected from SEQ ID NO: 156, SEQ ID NO: 157,
and SEQ ID NO: 158; [0878] o) 1, 2, or 3 heavy chain CDR sequences
selected from SEQ ID NO: 166, SEQ ID NO: 167, and SEQ ID NO: 168;
[0879] p) 1, 2, or 3 heavy chain CDR sequences selected from SEQ ID
NO: 176, SEQ ID NO: 177, and SEQ ID NO: 178; [0880] q) 1, 2, or 3
heavy chain CDR sequences selected from SEQ ID NO: 186, SEQ ID NO:
187, and SEQ ID NO: 188; [0881] r) 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 196, SEQ ID NO: 197, and SEQ ID
NO: 198; [0882] s) 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 206, SEQ ID NO: 207, and SEQ ID NO: 208; [0883] t)
1, 2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 216,
SEQ ID NO: 217, and SEQ ID NO: 218; [0884] u) 1, 2, or 3 heavy
chain CDR sequences selected from SEQ ID NO: 226, SEQ ID NO: 227,
and SEQ ID NO: 228; [0885] v) 1, 2, or 3 heavy chain CDR sequences
selected from SEQ ID NO: 236, SEQ ID NO: 237, and SEQ ID NO: 238;
[0886] w) 1, 2, or 3 heavy chain CDR sequences selected from SEQ ID
NO: 246, SEQ ID NO: 247, and SEQ ID NO: 248; [0887] x) 1, 2, or 3
heavy chain CDR sequences selected from SEQ ID NO: 256, SEQ ID NO:
257, and SEQ ID NO: 258; [0888] y) 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 266, SEQ ID NO: 267, and SEQ ID
NO: 268; [0889] z) 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 276, SEQ ID NO: 277, and SEQ ID NO: 278; [0890] aa)
1, 2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 286,
SEQ ID NO: 287, and SEQ ID NO: 288; [0891] bb) 1, 2, or 3 heavy
chain CDR sequences selected from SEQ ID NO: 296, SEQ ID NO: 297,
and SEQ ID NO: 298; [0892] cc) 1, 2, or 3 heavy chain CDR sequences
selected from SEQ ID NO: 306, SEQ ID NO: 307, and SEQ ID NO: 308;
[0893] dd) 1, 2, or 3 heavy chain CDR sequences selected from SEQ
ID NO: 316, SEQ ID NO: 317, and SEQ ID NO: 318; [0894] ee) 1, 2, or
3 heavy chain CDR sequences selected from SEQ ID NO: 326, SEQ ID
NO: 327, and SEQ ID NO: 328; [0895] ff) 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 336, SEQ ID NO: 337, and SEQ ID
NO: 338; [0896] gg) 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 346, SEQ ID NO: 347, and SEQ ID NO: 348; [0897] hh)
1, 2, or 3 heavy chain CDR sequences selected from SEQ ID NO: 356,
SEQ ID NO: 357, and SEQ ID NO: 358; [0898] ii) 1, 2, or 3 heavy
chain CDR sequences selected from SEQ ID NO: 366, SEQ ID NO: 367,
and SEQ ID NO: 368; [0899] jj) 1, 2, or 3 heavy chain CDR sequences
selected from SEQ ID NO: 376, SEQ ID NO: 377, and SEQ ID NO: 378;
[0900] kk) 1, 2, or 3 heavy chain CDR sequences selected from SEQ
ID NO: 386, SEQ ID NO: 387, and SEQ ID NO: 388; [0901] ll) 1, 2, or
3 heavy chain CDR sequences selected from SEQ ID NO: 396, SEQ ID
NO: 397, and SEQ ID NO: 398; [0902] mm) 1, 2, or 3 heavy chain CDR
sequences selected from SEQ ID NO: 406, SEQ ID NO: 407, and SEQ ID
NO: 408; [0903] nn) 1, 2, or 3 heavy chain CDR sequences selected
from SEQ ID NO: 416, SEQ ID NO: 417, and SEQ ID NO: 418; or [0904]
oo) 1, 2, or 3 heavy chain CDR sequences selected from SEQ ID NO:
426, SEQ ID NO: 427, and SEQ ID NO: 428. [0905] 4. The antibody or
antigen binding fragment of any of the preceding embodiments,
comprising: [0906] a) 1, 2, or 3 light chain CDR sequences selected
from SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6; [0907] b) 1, 2,
or 3 light chain CDR sequences selected from SEQ ID NO: 14, SEQ ID
NO: 15, and SEQ ID NO: 16; [0908] c) 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 24, SEQ ID NO: 25 and SEQ ID NO:
26; [0909] d) 1, 2, or 3 light chain CDR sequences selected from
SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36; [0910] e) 1, 2, or
3 light chain CDR sequences selected from SEQ ID NO: 44, SEQ ID NO:
45 and SEQ ID NO: 46; [0911] f) 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 54, SEQ ID NO: 55 and SEQ ID NO:
56; [0912] g) 1, 2, or 3 light chain CDR sequences selected from
SEQ ID NO: 68, SEQ ID NO: 69, and SEQ ID NO: 70; [0913] h) 1, 2, or
3 light chain CDR sequences selected from SEQ ID NO: 78, SEQ ID NO:
79, and SEQ ID NO: 80; and [0914] i) 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 88, SEQ ID NO: 89, and SEQ ID
NO: 90. [0915] j) 1, 2, or 3 light chain CDR sequences selected
from SEQ ID NO: 98, SEQ ID NO: 99, and SEQ ID NO: 100; [0916] k) 1,
2, or 3 light chain CDR sequences selected from SEQ ID NO: 108, SEQ
ID NO: 109, and SEQ ID NO: 110; [0917] l) 1, 2, or 3 light chain
CDR sequences selected from SEQ ID NO: 139, SEQ ID NO: 140, and SEQ
ID NO: 141; [0918] m) 1, 2, or 3 light chain CDR sequences selected
from SEQ ID NO: 149, SEQ ID NO: 150, and SEQ ID NO: 151; [0919] n)
1, 2, or 3 light chain CDR sequences selected from SEQ ID NO: 159,
SEQ ID NO: 160, and SEQ ID NO: 161; [0920] o) 1, 2, or 3 light
chain CDR sequences selected from SEQ ID NO: 169, SEQ ID NO: 170,
and SEQ ID NO: 171; [0921] p) 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 179, SEQ ID NO: 180, and SEQ ID NO: 181;
[0922] q) 1, 2, or 3 light chain CDR sequences selected from SEQ ID
NO: 189, SEQ ID NO: 190, and SEQ ID NO: 191; [0923] r) 1, 2, or 3
light chain CDR sequences selected from SEQ ID NO: 199, SEQ ID NO:
200, and SEQ ID NO: 201; [0924] s) 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 209, SEQ ID NO: 210, and SEQ ID
NO: 211; [0925] t) 1, 2, or 3 light chain CDR sequences selected
from SEQ ID NO: 219, SEQ ID NO: 220, and SEQ ID NO: 221; [0926] u)
1, 2, or 3 light chain CDR sequences selected from SEQ ID NO: 229,
SEQ ID NO: 230, and SEQ ID NO: 231; [0927] v) 1, 2, or 3 light
chain CDR sequences selected from SEQ ID NO: 239, SEQ ID NO: 240,
and SEQ ID NO: 241; [0928] w) 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 249, SEQ ID NO: 250, and SEQ ID NO: 251;
[0929] x) 1, 2, or 3 light chain CDR sequences selected from SEQ ID
NO: 259, SEQ ID NO: 260, and SEQ ID NO: 261; [0930] y) 1, 2, or 3
light chain CDR sequences selected from SEQ ID NO: 269, SEQ ID NO:
270, and SEQ ID NO: 271; [0931] z) 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 279, SEQ ID NO: 280, and SEQ ID
NO: 281; [0932] aa) 1, 2, or 3 light chain CDR sequences selected
from SEQ ID NO: 289, SEQ ID NO: 290, and SEQ ID NO: 291; [0933] bb)
1, 2, or 3 light chain CDR sequences selected from SEQ ID NO: 299,
SEQ ID NO: 300, and SEQ ID NO: 301; [0934] cc) 1, 2, or 3 light
chain CDR sequences selected from SEQ ID NO: 309, SEQ ID NO: 310,
and SEQ ID NO: 311; [0935] dd) 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 319, SEQ ID NO: 320, and SEQ ID NO: 321;
[0936] ee) 1, 2, or 3 light chain CDR sequences selected from SEQ
ID NO: 329, SEQ ID NO: 330, and SEQ ID NO: 331; [0937] ff) 1, 2, or
3 light chain CDR sequences selected from SEQ ID NO: 339, SEQ ID
NO: 340, and SEQ ID NO: 341; [0938] gg) 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 349, SEQ ID NO: 350, and SEQ ID
NO: 351; [0939] hh) 1, 2, or 3 light chain CDR sequences selected
from SEQ ID NO: 359, SEQ ID NO: 360, and SEQ ID NO: 361; [0940] ii)
1, 2, or 3 light chain CDR sequences selected from SEQ ID NO: 369,
SEQ ID NO: 370, and SEQ ID NO: 371; [0941] jj) 1, 2, or 3 light
chain CDR sequences selected from SEQ ID NO: 379, SEQ ID NO: 380,
and SEQ ID NO: 381; [0942] kk) 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 389, SEQ ID NO: 390, and SEQ ID NO: 391;
[0943] ll) 1, 2, or 3 light chain CDR sequences selected from SEQ
ID NO: 399, SEQ ID NO: 400, and SEQ ID NO: 401; [0944] mm) 1, 2, or
3 light chain CDR sequences selected from SEQ ID NO: 409, SEQ ID
NO: 410, and SEQ ID NO: 411; [0945] nn) 1, 2, or 3 light chain CDR
sequences selected from SEQ ID NO: 419, SEQ ID NO: 420, and SEQ ID
NO: 421; or [0946] oo) 1, 2, or 3 light chain CDR sequences
selected from SEQ ID NO: 429, SEQ ID NO: 430, and SEQ ID NO: 431.
[0947] 5. The antibody or antigen binding fragment of any of the
preceding embodiments, comprising: [0948] a) a heavy chain CDR1
(HCDR1) comprising the sequence of SEQ ID NO: 1, a heavy chain CDR2
(HCDR2) comprising the sequence of SEQ ID NO: 2, a heavy chain CDR3
(HCDR3) comprising the sequence of SEQ ID NO: 3; a light chain CDR1
(LCDR1) comprising the sequence of SEQ ID NO: 4, a light chain CDR2
(LCDR2) comprising the sequence of SEQ ID NO: 5, and a light chain
CDR3 (LCDR3) comprising the sequence of SEQ ID NO: 6; [0949] b) a
HCDR1 comprising the sequence of SEQ ID NO: 11, a HCDR2 comprising
the sequence of SEQ ID NO: 12, a HCDR3 comprising the sequence of
SEQ ID NO: 13, a LCDR1 comprising the sequence of SEQ ID NO: 14, a
LCDR2 comprising the sequence of SEQ ID NO: 15, and a LCDR3
comprising the sequence of SEQ ID NO: 16; [0950] c) a HCDR1
comprising the sequence of SEQ ID NO: 21, a HCDR2 comprising the
sequence of SEQ ID NO: 22, a HCDR3 comprising the sequence of SEQ
ID NO: 23, a LCDR1 comprising the sequence of SEQ ID NO: 24, a
LCDR2 comprising the sequence of SEQ ID NO: 25, and a LCDR3
comprising the sequence of SEQ ID NO: 26; [0951] d) a HCDR1
comprising the sequence of SEQ ID NO: 31, a HCDR2 comprising the
sequence of SEQ ID NO: 32, a HCDR3 comprising the sequence of SEQ
ID NO: 33, a LCDR1 comprising the sequence of SEQ ID NO: 34, a
LCDR2 comprising the sequence of SEQ ID NO: 35, and a LCDR3
comprising the sequence of SEQ ID NO: 36; [0952] e) a HCDR1
comprising the sequence of SEQ ID NO: 41, a HCDR2 comprising the
sequence of SEQ ID NO: 42, a HCDR3 comprising the sequence of SEQ
ID NO: 43, a LCDR1 comprising the sequence of SEQ ID NO: 44, a
LCDR2 comprising the sequence of SEQ ID NO: 45, and a LCDR3
comprising the sequence of SEQ ID NO: 46; [0953] f) a HCDR1
comprising the sequence of SEQ ID NO: 51, a HCDR2 comprising the
sequence of SEQ ID NO: 52, a HCDR3 comprising the sequence of SEQ
ID NO: 53, a LCDR1 comprising the sequence of SEQ ID NO: 54, a
LCDR2 comprising the sequence of SEQ ID NO: 55, and a LCDR3
comprising the sequence of SEQ ID NO: 56; [0954] g) a HCDR1
comprising the sequence of SEQ ID NO: 65, a HCDR2 comprising the
sequence of SEQ ID NO: 66, a HCDR3 comprising the sequence of SEQ
ID NO: 67, a LCDR1 comprising the sequence of SEQ ID NO: 68, a
LCDR2 comprising the sequence of SEQ ID NO: 69, and a LCDR3
comprising the sequence of SEQ ID NO: 70; [0955] h) a HCDR1
comprising the sequence of SEQ ID NO: 75, a HCDR2 comprising the
sequence of SEQ ID NO: 76, a HCDR3 comprising the sequence of SEQ
ID NO: 77, a LCDR1 comprising the sequence of SEQ ID NO: 78, a
LCDR2 comprising the sequence of SEQ ID NO: 79, and a LCDR3
comprising the sequence of SEQ ID NO: 80; [0956] i) a HCDR1
comprising the sequence of SEQ ID NO: 85, a HCDR2 comprising the
sequence of SEQ ID NO: 86, a HCDR3 comprising the sequence of SEQ
ID NO: 87, a LCDR1 comprising the sequence of SEQ ID NO: 88, a
LCDR2 comprising the sequence of SEQ ID NO: 89, and a LCDR3
comprising the sequence of SEQ ID NO: 90; [0957] j) a HCDR1
comprising the sequence of SEQ ID NO: 95, a HCDR2 comprising the
sequence of SEQ ID NO: 96, a HCDR3 comprising the sequence of SEQ
ID NO: 97, a LCDR1 comprising the sequence of SEQ ID NO: 98, a
LCDR2 comprising the sequence of SEQ ID NO: 99, and a LCDR3
comprising the sequence of SEQ ID NO: 100; [0958] k) a HCDR1
comprising the sequence of SEQ ID NO: 105, a HCDR2 comprising the
sequence of SEQ ID NO: 106, a HCDR3 comprising the sequence of SEQ
ID NO: 107, a LCDR1 comprising the sequence of SEQ ID NO: 108, a
LCDR2 comprising the sequence of SEQ ID NO: 109, and a LCDR3
comprising the sequence of SEQ ID NO: 110; [0959] l) a HCDR1
comprising the sequence of SEQ ID NO: 136, a HCDR2 comprising the
sequence of SEQ ID NO: 137, a HCDR3 comprising the sequence of SEQ
ID NO: 138, a LCDR1 comprising the sequence of SEQ ID NO: 139, a
LCDR2 comprising the sequence of SEQ ID NO: 140, and a LCDR3
comprising the sequence of SEQ ID NO: 141; [0960] m) HCDR1
comprising the sequence of SEQ ID NO: 146, a HCDR2 comprising the
sequence of SEQ ID NO: 147, a HCDR3 comprising the sequence of SEQ
ID NO: 148, a LCDR1 comprising the sequence of SEQ ID NO: 149, a
LCDR2 comprising the sequence of SEQ ID NO: 150, and a LCDR3
comprising the sequence of SEQ ID NO: 151; [0961] n) HCDR1
comprising the sequence of SEQ ID NO: 156, a HCDR2 comprising the
sequence of SEQ ID NO: 157, a HCDR3 comprising the sequence of SEQ
ID NO: 158, a LCDR1 comprising the sequence of SEQ ID NO: 159, a
LCDR2 comprising the sequence of SEQ ID NO: 160, and a LCDR3
comprising the sequence of SEQ ID NO: 161; [0962] o) HCDR1
comprising the sequence of SEQ ID NO: 166, a HCDR2 comprising the
sequence of SEQ ID NO: 167, a HCDR3 comprising the sequence of SEQ
ID NO: 168, a LCDR1 comprising the sequence of SEQ ID NO: 169, a
LCDR2 comprising the sequence of SEQ ID NO: 170, and a LCDR3
comprising the sequence of SEQ ID NO: 171; [0963] p) HCDR1
comprising the sequence of SEQ ID NO: 176, a HCDR2 comprising the
sequence of SEQ ID NO: 177, a HCDR3 comprising the sequence of SEQ
ID NO: 178, a LCDR1 comprising the sequence of SEQ ID NO: 179, a
LCDR2 comprising the sequence of SEQ ID NO: 180, and a LCDR3
comprising the sequence of SEQ ID NO: 181; [0964] q) HCDR1
comprising the sequence of SEQ ID NO: 186, a HCDR2 comprising the
sequence of SEQ ID NO: 187, a HCDR3 comprising the sequence of SEQ
ID NO: 188, a LCDR1 comprising the sequence of SEQ ID NO: 189, a
LCDR2 comprising the sequence of SEQ ID NO: 190, and a LCDR3
comprising the sequence of SEQ ID NO: 191;
[0965] r) HCDR1 comprising the sequence of SEQ ID NO: 196, a HCDR2
comprising the sequence of SEQ ID NO: 197, a HCDR3 comprising the
sequence of SEQ ID NO: 198, a LCDR1 comprising the sequence of SEQ
ID NO: 199, a LCDR2 comprising the sequence of SEQ ID NO: 200, and
a LCDR3 comprising the sequence of SEQ ID NO: 201; [0966] s) HCDR1
comprising the sequence of SEQ ID NO: 206, a HCDR2 comprising the
sequence of SEQ ID NO: 207, a HCDR3 comprising the sequence of SEQ
ID NO: 208, a LCDR1 comprising the sequence of SEQ ID NO: 209, a
LCDR2 comprising the sequence of SEQ ID NO: 210, and a LCDR3
comprising the sequence of SEQ ID NO: 211; [0967] t) HCDR1
comprising the sequence of SEQ ID NO: 216, a HCDR2 comprising the
sequence of SEQ ID NO: 217, a HCDR3 comprising the sequence of SEQ
ID NO: 218, a LCDR1 comprising the sequence of SEQ ID NO: 219, a
LCDR2 comprising the sequence of SEQ ID NO: 220, and a LCDR3
comprising the sequence of SEQ ID NO: 221; [0968] u) HCDR1
comprising the sequence of SEQ ID NO: 226, a HCDR2 comprising the
sequence of SEQ ID NO: 227, a HCDR3 comprising the sequence of SEQ
ID NO: 228, a LCDR1 comprising the sequence of SEQ ID NO: 229, a
LCDR2 comprising the sequence of SEQ ID NO: 230, and a LCDR3
comprising the sequence of SEQ ID NO: 231; [0969] v) HCDR1
comprising the sequence of SEQ ID NO: 236, a HCDR2 comprising the
sequence of SEQ ID NO: 237, a HCDR3 comprising the sequence of SEQ
ID NO: 238, a LCDR1 comprising the sequence of SEQ ID NO: 239, a
LCDR2 comprising the sequence of SEQ ID NO: 240, and a LCDR3
comprising the sequence of SEQ ID NO: 241; [0970] w) HCDR1
comprising the sequence of SEQ ID NO: 246, a HCDR2 comprising the
sequence of SEQ ID NO: 247, a HCDR3 comprising the sequence of SEQ
ID NO: 248, a LCDR1 comprising the sequence of SEQ ID NO: 249, a
LCDR2 comprising the sequence of SEQ ID NO: 250, and a LCDR3
comprising the sequence of SEQ ID NO: 251; [0971] x) HCDR1
comprising the sequence of SEQ ID NO: 256, a HCDR2 comprising the
sequence of SEQ ID NO: 257, a HCDR3 comprising the sequence of SEQ
ID NO: 258, a LCDR1 comprising the sequence of SEQ ID NO: 259, a
LCDR2 comprising the sequence of SEQ ID NO: 260, and a LCDR3
comprising the sequence of SEQ ID NO: 261; [0972] y) HCDR1
comprising the sequence of SEQ ID NO: 266, a HCDR2 comprising the
sequence of SEQ ID NO: 267, a HCDR3 comprising the sequence of SEQ
ID NO: 268, a LCDR1 comprising the sequence of SEQ ID NO: 269, a
LCDR2 comprising the sequence of SEQ ID NO: 270, and a LCDR3
comprising the sequence of SEQ ID NO: 271; [0973] z) HCDR1
comprising the sequence of SEQ ID NO: 276, a HCDR2 comprising the
sequence of SEQ ID NO: 277, a HCDR3 comprising the sequence of SEQ
ID NO: 278, a LCDR1 comprising the sequence of SEQ ID NO: 279, a
LCDR2 comprising the sequence of SEQ ID NO: 280, a LCDR3 comprising
the sequence of SEQ ID NO: 281; [0974] aa) HCDR1 comprising the
sequence of SEQ ID NO: 286, a HCDR2 comprising the sequence of SEQ
ID NO: 287, a HCDR3 comprising the sequence of SEQ ID NO: 288, a
LCDR1 comprising the sequence of SEQ ID NO: 289, a LCDR2 comprising
the sequence of SEQ ID NO: 290, a LCDR3 comprising the sequence of
SEQ ID NO: 291; [0975] bb) HCDR1 comprising the sequence of SEQ ID
NO: 296, a HCDR2 comprising the sequence of SEQ ID NO: 297, a HCDR3
comprising the sequence of SEQ ID NO: 298, a LCDR1 comprising the
sequence of SEQ ID NO: 299, a LCDR2 comprising the sequence of SEQ
ID NO: 300, a LCDR3 comprising the sequence of SEQ ID NO: 301;
[0976] cc) HCDR1 comprising the sequence of SEQ ID NO: 306, a HCDR2
comprising the sequence of SEQ ID NO: 307, a HCDR3 comprising the
sequence of SEQ ID NO: 308, a LCDR1 comprising the sequence of SEQ
ID NO: 309, a LCDR2 comprising the sequence of SEQ ID NO: 310, a
LCDR3 comprising the sequence of SEQ ID NO: 311; [0977] dd) HCDR1
comprising the sequence of SEQ ID NO: 316, a HCDR2 comprising the
sequence of SEQ ID NO: 317, a HCDR3 comprising the sequence of SEQ
ID NO: 318, a LCDR1 comprising the sequence of SEQ ID NO: 319, a
LCDR2 comprising the sequence of SEQ ID NO: 320, a LCDR3 comprising
the sequence of SEQ ID NO: 321; [0978] ee) HCDR1 comprising the
sequence of SEQ ID NO: 326, a HCDR2 comprising the sequence of SEQ
ID NO: 327, a HCDR3 comprising the sequence of SEQ ID NO: 328, a
LCDR1 comprising the sequence of SEQ ID NO: 329, a LCDR2 comprising
the sequence of SEQ ID NO: 330, a LCDR3 comprising the sequence of
SEQ ID NO: 331; [0979] ff) HCDR1 comprising the sequence of SEQ ID
NO: 336, a HCDR2 comprising the sequence of SEQ ID NO: 337, a HCDR3
comprising the sequence of SEQ ID NO: 338, a LCDR1 comprising the
sequence of SEQ ID NO: 339, a LCDR2 comprising the sequence of SEQ
ID NO: 340, a LCDR3 comprising the sequence of SEQ ID NO: 341;
[0980] gg) HCDR1 comprising the sequence of SEQ ID NO: 346, a HCDR2
comprising the sequence of SEQ ID NO: 347, a HCDR3 comprising the
sequence of SEQ ID NO: 348, a LCDR1 comprising the sequence of SEQ
ID NO: 349, a LCDR2 comprising the sequence of SEQ ID NO: 350, a
LCDR3 comprising the sequence of SEQ ID NO: 351; [0981] hh) HCDR1
comprising the sequence of SEQ ID NO: 356, a HCDR2 comprising the
sequence of SEQ ID NO: 357, a HCDR3 comprising the sequence of SEQ
ID NO: 358, a LCDR1 comprising the sequence of SEQ ID NO: 359, a
LCDR2 comprising the sequence of SEQ ID NO: 360, a LCDR3 comprising
the sequence of SEQ ID NO: 361; [0982] ii) HCDR1 comprising the
sequence of SEQ ID NO: 366, a HCDR2 comprising the sequence of SEQ
ID NO: 367, a HCDR3 comprising the sequence of SEQ ID NO: 368, a
LCDR1 comprising the sequence of SEQ ID NO: 369, a LCDR2 comprising
the sequence of SEQ ID NO: 370, a LCDR3 comprising the sequence of
SEQ ID NO: 371; [0983] jj) HCDR1 comprising the sequence of SEQ ID
NO: 376, a HCDR2 comprising the sequence of SEQ ID NO: 377, a HCDR3
comprising the sequence of SEQ ID NO: 378, a LCDR1 comprising the
sequence of SEQ ID NO: 379, a LCDR2 comprising the sequence of SEQ
ID NO: 380, a LCDR3 comprising the sequence of SEQ ID NO: 381;
[0984] kk) HCDR1 comprising the sequence of SEQ ID NO: 386, a HCDR2
comprising the sequence of SEQ ID NO: 387, a HCDR3 comprising the
sequence of SEQ ID NO: 388, a LCDR1 comprising the sequence of SEQ
ID NO: 389, a LCDR2 comprising the sequence of SEQ ID NO: 390, a
LCDR3 comprising the sequence of SEQ ID NO: 391; [0985] ll) HCDR1
comprising the sequence of SEQ ID NO: 396, a HCDR2 comprising the
sequence of SEQ ID NO: 397, a HCDR3 comprising the sequence of SEQ
ID NO: 398, a LCDR1 comprising the sequence of SEQ ID NO: 399, a
LCDR2 comprising the sequence of SEQ ID NO: 400, a LCDR3 comprising
the sequence of SEQ ID NO: 401; [0986] mm) HCDR1 comprising the
sequence of SEQ ID NO: 406, a HCDR2 comprising the sequence of SEQ
ID NO: 407, a HCDR3 comprising the sequence of SEQ ID NO: 408, a
LCDR1 comprising the sequence of SEQ ID NO: 409, a LCDR2 comprising
the sequence of SEQ ID NO: 410, a LCDR3 comprising the sequence of
SEQ ID NO: 411; [0987] nn) HCDR1 comprising the sequence of SEQ ID
NO: 416, a HCDR2 comprising the sequence of SEQ ID NO: 417, a HCDR3
comprising the sequence of SEQ ID NO: 418, a LCDR1 comprising the
sequence of SEQ ID NO: 419, a LCDR2 comprising the sequence of SEQ
ID NO: 420, a LCDR3 comprising the sequence of SEQ ID NO: 421; or
[0988] oo) HCDR1 comprising the sequence of SEQ ID NO: 426, a HCDR2
comprising the sequence of SEQ ID NO: 427, a HCDR3 comprising the
sequence of SEQ ID NO: 428, a LCDR1 comprising the sequence of SEQ
ID NO: 429, a LCDR2 comprising the sequence of SEQ ID NO: 430, a
LCDR3 comprising the sequence of SEQ ID NO: 431. [0989] 6. The
antibody or antigen binding fragment of any of the preceding
embodiments, comprising a pair of heavy chain variable region and
light chain variable region sequences selected from the group
consisting of: SEQ ID NOs: 7/8, 17/18, 27/28, 37/38, 47/48, 57/58,
61/62, 71/72, 81/82, 91/92, 101/102, 111/112, 142/143, 152/153,
162/163, 172/173, 182/183, 192/193, 202/203, 212/213, 222/223,
232/233, 242/243, 252/253, 262/263, 272/273, 282/283, 292/293,
302/303, 312/313, 322/323, 332/333, 342/343, 352/353, 362/363,
372/373, 382/383, 392/393, 402/403, 412/413, 422/423 and 432/433,
or a pair of homologous sequences thereof having at least 80%
sequence identity yet retaining binding specificity to RBD of spike
protein of SARS-CoV-2. [0990] 7. The antibody or antigen binding
fragment of embodiments 1 or 2, which is a variant of antibody
P2A-1A8, P2A-1A9, P2B-2G11, P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4,
P2C-1A3, P2C-1C8, P2C-1C10, P2C-1D5, P2C-1F11, P2B-1G5, P2B-1A1,
P2C-1D7, P2B-1A10, P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7,
P5A-3C8, P5A-1D2, P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6,
P4B-1F4, P5A-1B6, P5A-1B8, P5A-1B9, P5A-1D1, P5A-1D10, P5A-2D11,
P5A-2G9, P5A-2H3, P5A-3A1, P5A-3A6, P5A-3B4, P5A-3C12, or P22A-1D1,
which comprises: [0991] a) a heavy chain CDR1 (HCDR1) sequence
having at least 80% (e.g. at least 85%, 88%, 90%, 91%, 92%, 93%,
94%, 95%, 96%, 97%, 98%, 99%) sequence identity to a HCDR1 sequence
of the parent antibody listed in Table 1, and/or [0992] b) a heavy
chain CDR2 (HCDR2) sequence having at least 80% (e.g. at least 85%,
88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) sequence
identity to a HCDR2 sequence of the parent antibody listed in Table
1, and/or [0993] c) a heavy chain CDR3 (HCDR3) sequence having at
least 80% (e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%,
96%, 97%, 98%, 99%) sequence identity to a HCDR3 sequence of the
parent antibody listed in Table 1, and/or [0994] d) a light chain
CDR1 (LCDR1) sequence having at least 80% (e.g. at least 85%, 88%,
90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) sequence identity
to a LCDR1 sequence of the parent antibody listed in Table 1,
and/or [0995] e) a light chain CDR2 (LCDR2) sequence having at
least 80% (e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%,
96%, 97%, 98%, 99%) sequence identity to a LCDR2 sequence of the
parent antibody listed in Table 1, and/or [0996] f) a light chain
CDR3 (LCDR3) sequence having at least 80% (e.g. at least 85%, 88%,
90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) sequence identity
to a LCDR3 sequence of the parent antibody listed in Table 1, and
[0997] which retains the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than its parent antibody. [0998] 8. The antibody
or antigen binding fragment of embodiment 7, which comprises an
HCDR1 having no more than 3, 2, or 1 amino acid mutations in a
HCDR1 sequence of the parent antibody listed in Table 1, an HCDR2
having no more than 6, 5, 4, 3, 2, or 1 amino acid mutations in a
HCDR2 sequence of the parent antibody listed in Table 1, HCDR3
having no more than 6, 5, 4, 3, 2, or 1 amino acid mutations in a
HCDR3 sequence of the parent antibody listed in Table 1, LCDR1
having no more than 2 or 1 amino acid mutations in a LCDR1 sequence
of the parent antibody listed in Table 1, LCDR2 having no more than
3, 2, or 1 amino acid mutations in a LCDR2 sequence of the parent
antibody listed in Table 1, and/or LCDR3 having no more than 3, 2,
or 1 amino acid mutations in a LCDR3 sequence of the parent
antibody listed in Table 1. [0999] 9. The antibody or antigen
binding fragment of any of the preceding embodiments, which
comprises: [1000] a) at least one heavy chain CDR sequence having
no more than 3, 2, or 1 amino acid substitutions in a heavy chain
CDR sequence of the parent antibody listed in Table 1, or [1001] b)
at least two heavy chain CDR sequences each having no more than 3,
2, or 1 amino acid substitutions in a heavy chain CDR sequence of
the parent antibody listed in Table 1, or [1002] c) three heavy
chain CDR sequences each having no more than 3, 2, or 1 amino acid
substitutions in a heavy chain CDR sequence of the parent antibody
listed in Table 1, or [1003] d) at least one light chain sequence
having no more than 3, 2, or 1 amino acid substitutions in a heavy
chain CDR sequence of the parent antibody listed in Table 1, or
[1004] e) at least two light chain CDR sequences each having no
more than 3, 2, or 1 amino acid substitutions in a heavy chain CDR
sequence of the parent antibody listed in Table 1, or [1005] f)
three light chain CDR sequences each having no more than 3, 2, or 1
amino acid substitutions in a heavy chain CDR sequence of the
parent antibody listed in Table 1. [1006] 10. The antibody or
antigen binding fragment of embodiments 1 or 2, which is a variant
of antibody P2B-2F6 and comprises: [1007] a) a heavy chain CDR1
(HCDR1) sequence having at least 80% (e.g. at least 85%, 88%, 90%,
91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) sequence identity to
SEQ ID NO: 41, and/or [1008] b) a heavy chain CDR2 (HCDR2) sequence
having at least 80% (e.g. at least 85%, 88%, 90%, 91%, 92%, 93%,
94%, 95%, 96%, 97%, 98%, 99%) sequence identity to SEQ ID NO: 42,
and/or [1009] c) a heavy chain CDR3 (HCDR3) sequence having at
least 80% (e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%,
96%, 97%, 98%, 99%) sequence identity to SEQ ID NO: 43, and/or
[1010] d) a light chain CDR1 (LCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 44, and/or [1011] e) a
light chain CDR2 (LCDR2) sequence having at least 80% (e.g. at
least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%)
sequence identity to SEQ ID NO: 45, and/or [1012] f) a light chain
CDR3 (LCDR3) sequence having at least 80% (e.g. at least 85%, 88%,
90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) sequence identity
to SEQ ID NO: 46, and [1013] which retains the binding specificity
to SARS-COV-2, optionally having binding affinity to SARS-COV-2 at
a level similar to or even higher than antibody P2B-2F6. [1014] 11.
The antibody or antigen binding fragment of embodiment 10, which
comprises an HCDR1 having no more than 4, 3, 2, or 1 amino acid
mutations in SEQ ID NO: 41, an HCDR2 having no more than 3, 2, or 1
amino acid mutations in SEQ ID NO: 42, HCDR3 having no more than 6,
5, 4, 3, 2, or 1 amino acid substitutions in SEQ ID NO: 43, LCDR1
having no more than 4, 3, 2, or 1 amino acid mutations in SEQ ID
NO: 44, LCDR2 having no more than 3, 2, or 1 amino acid mutations
in SEQ ID NO: 45, and/or LCDR3 having no more than 4, 3, 2, or 1
amino acid mutations in SEQ ID NO: 46. [1015] 12. The antibody or
antigen binding fragment of embodiments 10 or 11, which retains the
entirety of or at least part of the paratope of antibody P2B-2F6
while one or more of the amino acid residues outside the paratope
of the antibody may be mutated. [1016] 13. The antibody or antigen
binding fragment of embodiment 12, wherein the paratope of antibody
P2B-2F6 comprises or consists of: Y27, S28, S30, S31, and Y33 of
HCDR1, H54 of HCDR2, G102, I103, V105, V106 and P107 of HCDR3, and
G31, Y32 and N33 of LCDR1, wherein the numbering of residues in the
heavy chain CDRs is according to SEQ ID NO: 47, and the numbering
of residues in the light chain CDR is according to SEQ ID NO:
48.
[1017] 14. The antibody or antigen binding fragment of embodiments
1 or 2, which is a variant of antibody P2C-1F11, which comprises:
[1018] a) a heavy chain CDR1 (HCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 105, and/or [1019] b) a
heavy chain CDR2 (HCDR2) sequence having at least 80% (e.g. at
least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%)
sequence identity to SEQ ID NO: 106, and/or [1020] c) a heavy chain
CDR3 (HCDR3) sequence having at least 80% (e.g. at least 85%, 88%,
90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) sequence identity
to SEQ ID NO: 107, and/or [1021] d) a light chain CDR1 (LCDR1)
sequence having at least 80% (e.g. at least 85%, 88%, 90%, 91%,
92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%) sequence identity to SEQ ID
NO: 108, and/or [1022] e) a light chain CDR2 (LCDR2) sequence
having at least 80% (e.g. at least 85%, 88%, 90%, 91%, 92%, 93%,
94%, 95%, 96%, 97%, 98%, 99%) sequence identity to SEQ ID NO: 109,
and/or [1023] f) a light chain CDR3 (LCDR3) sequence having at
least 80% (e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%,
96%, 97%, 98%, 99%) sequence identity to SEQ ID NO: 110, and [1024]
which retains the binding specificity to SARS-COV-2, optionally
having binding affinity to SARS-COV-2 at a level similar to or even
higher than antibody P2C-1F11. [1025] 15. The antibody or antigen
binding fragment of embodiment 14, which comprises an HCDR1 having
no more than 4, 3, 2, or 1 amino acid mutations in SEQ ID NO: 105,
an HCDR2 having no more than 3, 2, or 1 amino acid mutations in SEQ
ID NO: 106, HCDR3 having no more than 6, 5, 4, 3, 2, or 1 amino
acid mutations in SEQ ID NO: 107, LCDR1 having no more than 4, 3,
2, or 1 amino acid mutations in SEQ ID NO: 108, LCDR2 having no
more than 3, 2, or 1 amino acid mutations in SEQ ID NO: 109, and/or
LCDR3 having no more than 4, 3, 2, or 1 amino acid mutations in SEQ
ID NO: 110, and in the meantime retain the binding specificity to
SARS-COV-2, optionally having binding affinity to SARS-COV-2 at a
level similar to or even higher than antibody P2C-1F11. [1026] 16.
The antibody or antigen binding fragment of embodiment 14, which
retains the entirety of or at least part of the paratope of
antibody P2C-1F11 while one or more of the amino acid residues
outside the paratope of the antibody may be mutated. [1027] 17. The
antibody or antigen binding fragment of embodiment 16, wherein the
paratope of antibody P2C-1F11 comprises or consists of: G26, 127,
T28, S31, N32 and Y33 of HCDR1, Y52, S53, G54, and S56 of HCDR2,
R97, L99, V100, V101, Y102 and D105 of HCDR3, and S28, S30 and Y33
of LCDR1 of LCDR1, wherein the numbering of residues in heavy chain
is according to SEQ ID NO: 111, and the numbering of residues in
light chain CDR is according to SEQ ID NO: 112. [1028] 18. The
antibody or antigen binding fragment of embodiments 1 or 2, which
is a variant of antibody P22A-1D1, wherein the variant
comprises:
[1029] a) a heavy chain CDR1 (HCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 426, and/or
[1030] b) a heavy chain CDR2 (HCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 427, and/or
[1031] c) a heavy chain CDR3 (HCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 428, and/or
[1032] d) a light chain CDR1 (LCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 429, and/or
[1033] e) a light chain CDR2 (LCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 430, and/or
[1034] f) a light chain CDR3 (LCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 431, and
in the meantime retain the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than antibody P22A-1D1. [1035] 19. The antibody
or antigen binding fragment of embodiment 18, which comprises an
HCDR1 having no more than 6, 5, 4, 3, 2, or 1 amino acid mutations
in SEQ ID NO: 426, an HCDR2 having no more than 5, 4, 3, 2, or 1
amino acid mutations in SEQ ID NO: 427, an HCDR3 having no more
than 6, 5, 4, 3, 2, or 1 amino acid substitutions in SEQ ID NO:
428, an LCDR1 having no more than 5, 4, 3, 2, or 1 amino acid
mutations in SEQ ID NO: 429, LCDR2 having no more than 1 amino acid
mutations in SEQ ID NO: 430, and/or LCDR3 having no more than 4, 3,
2, or 1 amino acid mutations in SEQ ID NO: 431, and in the meantime
retain the binding specificity to SARS-COV-2, optionally having
binding affinity to SARS-COV-2 at a level similar to or even higher
than antibody P22A-1D1. [1036] 20. The antibody or antigen binding
fragment of embodiment 18, which retain the entirety of the
paratope of antibody P22A-1D1 while one or more of the amino acid
residues outside the paratope of the antibody may be mutated.
[1037] 21. The antibody or antigen binding fragment of embodiments
20, wherein the paratope of antibody P22A-1D1 comprises or consists
of: G26, F27, T28, S31, N32 and Y33 of HCDR1; Y52, S53, G54, and
S56 of HCDR2, Y58 of heavy chain framework region 3; R97, R99,
D100, Y101, Y102 and D105 of HCDR3; Q27, G28, 129, S30 and Y32 of
LCDR1; S67 of LCDR2; and/or H90, L91, N92 and Y94 of LCDR3; wherein
the numbering of residues in the heavy chain CDRs is according to
SEQ ID NO: 432, and the numbering of residues in the light chain
CDR is according to SEQ ID NO: 433. [1038] 22. The antibody or
antigen binding fragment of embodiments 1 or 2, which is a variants
of antibody P5A-1D2, wherein the variant comprises:
[1039] a) a heavy chain CDR1 (HCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 236, and/or
[1040] b) a heavy chain CDR2 (HCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 237, and/or
[1041] c) a heavy chain CDR3 (HCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 238, and/or
[1042] d) a light chain CDR1 (LCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 239, and/or
[1043] e) a light chain CDR2 (LCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 240, and/or
[1044] f) a light chain CDR3 (LCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 241, and
in the meantime retain the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than antibody P5A-1D2. [1045] 23. The antibody or
antigen binding fragment of embodiment 22, which comprises an HCDR1
having no more than 4, 3, 2, or 1 amino acid mutations in SEQ ID
NO: 236, an HCDR2 having no more than 3, 2, or 1 amino acid
mutations in SEQ ID NO: 237, HCDR3 having no more than 6, 5, 4, 3,
2, or 1 amino acid substitutions in SEQ ID NO: 238, LCDR1 having no
more than 4, 3, 2, or 1 amino acid mutations in SEQ ID NO: 239,
LCDR2 having no more than 3, 2, or 1 amino acid mutations in SEQ ID
NO: 240, and/or LCDR3 having no more than 4, 3, 2, or 1 amino acid
mutations in SEQ ID NO: 241, and in the meantime retain the binding
specificity to SARS-COV-2, optionally having binding affinity to
SARS-COV-2 at a level similar to or even higher than antibody
P5A-1D2. [1046] 24. The antibody or antigen binding fragment of
embodiment 22, which retain the entirety of the paratope of
antibody P5A-1D2 while one or more of the amino acid residues
outside the paratope of the antibody may be mutated. [1047] 25. The
antibody or antigen binding fragment of embodiment 24, wherein the
paratope of antibody P5A-1D2 comprises or consists of: G26, F27,
128, S31, N32 and Y33 of HCDR1; Y52, S53, G54, and S56 of HCDR2;
Y58 and R87 of heavy chain framework region 3, R97, L99, Q100,
V101, G102, A103, T104 and D106 of HCDR3; A31 and Y33 of LCDR1;
and/or S95 of LCDR3; wherein the numbering of residues in the heavy
chain CDRs is according to SEQ ID NO: 242, and the numbering of
residues in the light chain CDR is according to SEQ ID NO: 243.
[1048] 26. The antibody or antigen binding fragment of embodiments
1 or 2, which is a variants of antibody P5A-3C8, wherein the
variant comprises:
[1049] a) a heavy chain CDR1 (HCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 226, and/or
[1050] b) a heavy chain CDR2 (HCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 227, and/or
[1051] c) a heavy chain CDR3 (HCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 228, and/or
[1052] d) a light chain CDR1 (LCDR1) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 229, and/or
[1053] e) a light chain CDR2 (LCDR2) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 230, and/or
[1054] f) a light chain CDR3 (LCDR3) sequence having at least 80%
(e.g. at least 85%, 88%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,
98%, 99%) sequence identity to SEQ ID NO: 231, and
in the meantime retain the binding specificity to SARS-COV-2,
optionally having binding affinity to SARS-COV-2 at a level similar
to or even higher than antibody P5A-3C8. [1055] 27. The antibody or
antigen binding fragment of embodiment 26, which comprises an HCDR1
having no more than 4, 3, 2, or 1 amino acid mutations in SEQ ID
NO: 226, an HCDR2 having no more than 3, 2, or 1 amino acid
mutations in SEQ ID NO: 227, HCDR3 having no more than 6, 5, 4, 3,
2, or 1 amino acid substitutions in SEQ ID NO: 228, LCDR1 having no
more than 4, 3, 2, or 1 amino acid mutations in SEQ ID NO: 229,
LCDR2 having no more than 3, 2, or 1 amino acid mutations in SEQ ID
NO: 230, and/or LCDR3 having no more than 4, 3, 2, or 1 amino acid
mutations in SEQ ID NO: 231, and in the meantime retain the binding
specificity to SARS-COV-2, optionally having binding affinity to
SARS-COV-2 at a level similar to or even higher than antibody
P5A-3C8. [1056] 28. The antibody or antigen binding fragment of
embodiment 26, which retain the entirety of the paratope of
antibody P5A-3C8 while one or more of the amino acid residues
outside the paratope of the antibody may be mutated. [1057] 29. The
antibody or antigen binding fragment of embodiment 28, wherein the
paratope of antibody P5A-3C8 comprises or consists of: G26, F27,
T28, S31, N32 and Y33 of HCDR1; Y52, S53, G54, and S56 of HCDR2;
Y58 of heavy chain framework region 3, R97, L99, Q100, E101 and
H102 of HCDR3; and G28, 129, S30, S31 and Y32 of LCDR1; S67 of
LCDR2; G68 of light chain framework region 3, H90, L91, N92, S93
and Y94 of LCDR3; wherein the numbering of residues in the heavy
chain CDRs is according to SEQ ID NO: 232, and the numbering of
residues in the light chain CDR is according to SEQ ID NO: 233.
[1058] 30. The antibody or antigen binding fragment of any of the
preceding embodiments, further comprising an immunoglobulin
constant region, optionally a constant region of human
immunoglobulin, or optionally a constant region of human IgG.
[1059] 31. The antibody or antigen binding fragment of any of the
preceding embodiments, further comprising one or more amino acid
residue mutations yet retains binding specificity to SARS-CoV-2,
optionally binding affinity to RBD of spike protein of SARS-CoV-2.
[1060] 32. The antibody or antigen binding fragment of embodiment
31, which is an affinity variant, a glycosylation variant, a
cysteine-engineered variant, or an Fc variant. [1061] 33. The
antibody or antigen binding fragment of embodiment 32, wherein the
Fc variant comprises one or more amino acid residue modifications
or substitutions resulting in increased effector functions relative
to a wildtype Fc. [1062] 34. The antibody or antigen binding
fragment of embodiment 33, wherein the Fc variant comprises one or
more amino acid substitution(s) at one or more of the following
positions: 234, 235, 236, 238, 239, 240, 241, 243, 244, 245, 246,
247, 248, 249, 252, 254, 255, 256, 258, 260, 262, 263, 264, 265,
267, 268, 269, 270, 272, 274, 276, 278, 280, 283, 285, 286, 289,
290, 292, 293, 294, 295, 296, 298, 299, 300, 301, 303, 304, 305,
307, 309, 312, 313, 315, 320, 322, 324, 325, 326, 327, 329, 330,
331, 332, 333, 334, 335, 337, 338, 339, 340, 345, 360, 373, 376,
378, 382, 388, 389, 396, 398, 414, 416, 419, 430, 433, 434, 435,
436, 437, 438, 439 and 440 of the Fc region, wherein the numbering
of the residues in the Fc region is that of the EU index as in
Kabat. [1063] 35. The antibody or antigen binding fragment of
embodiment 34, wherein the Fc variant comprises one or more amino
acid substitution selected from the group consisting of 234Y, 235Q,
236A, 236W, 239D, 239E, 239M, 243L, 2471, 267E, 268D, 268E, 268F,
270E, 280H, 290S, 292P, 298A, 298D, 298V, 300L, 3051, 324T, 326A,
326D, 326W, 330L, 330M, 333S, 332D, 332E, 298A, 333A, 334A, 334E,
339D, 339Q, 345R, 396L, 430G, 440Y, or any combination thereof.
[1064] 36. The antibody or antigen binding fragment of embodiment
33, wherein the Fc variant comprises one or more amino acid residue
modifications or substitutions resulting in reduced effector
functions relative to a wildtype Fc. [1065] 37. The antibody or
antigen binding fragment of embodiment 36, wherein the Fc variant
comprises one or more amino acid substitution(s) at a position
selected from the group consisting of: 220, 226, 229, 233, 234,
235, 236, 237, 238, 267, 268, 269, 270, 297, 309, 318, 320, 322,
325, 328, 329, 330, and 331 of the Fc region, wherein the numbering
of the residues in the Fc region is that of the EU index as in
Kabat. [1066] 38. The antibody or antigen binding fragment of
embodiment 36, wherein the Fc variant comprises one or more amino
acid substitution(s) selected from the group consisting of 220S,
226S, 228P, 229S, 233P, 234V, 234G, 234A, 234F, 234A, 235A, 235G,
235E, 236E, 236R, 237A, 237K, 238S, 267R, 268A, 268Q, 269R, 297A,
297Q, 297G, 309L, 318A, 322A, 325L, 328R, 330S, 331S, and any
combination thereof. [1067] 39. The antibody or antigen binding
fragment of embodiment 33, wherein the Fc variant comprises one or
more amino acid residue modifications or substitutions resulting in
improved serum half-life or improved binding affinity to neonatal
Fc receptor (FcRn) at pH 6.0 while retaining minimal binding at pH
7.4. [1068] 40. The antibody or antigen binding fragment of
embodiment 39, wherein the Fc variant comprises one or more amino
acid substitution(s) at a position selected from the group
consisting of: 234, 235, 238, 250, 252, 254, 256; 259; 272, 305,
307, 308, 311, 312, 322, 328, 331, 378, 380, 382, 428, 432, 433,
434, 435, 436 and 437 (all positions by EU numbering). [1069] 41.
The antibody or antigen binding fragment of embodiment 40, wherein
the Fc variant comprises one or more amino acid substitution(s)
selected from the group consisting of 234F, 235Q, 238D, 250Q, 252T,
252Y, 254T, 256E, 2591, 272A, 305A, 307A, 308F, 311A, 322Q, 328E,
331S, 380A, 428L, 432C, 433K, 433S, 434S, 434Y, 434F, 434W, 434A,
435H, 436L, 437C and any combination thereof. [1070] 42. The
antibody or antigen binding fragment of embodiment 31, wherein at
least one of the substitutions or modifications is in one or more
of the CDR sequences, and/or in one or more of the non-CDR
sequences of the heavy chain variable region or light chain
variable region. [1071] 43. The antibody or an antigen-binding
fragment thereof of any one of the preceding embodiments, which is
a monoclonal antibody, a bispecific antibody, a multi-specific
antibody, a recombinant antibody, a chimeric antibody, a labeled
antibody, a bivalent antibody, an anti-idiotypic antibody, a fusion
protein, a dimerized or polymerized antibody, or a modified
antibody (e.g. glycosylated antibody). [1072] 44. The antibody or
antigen binding fragment of any of the preceding embodiments, which
is a diabody, a Fab, a Fab', a F(ab').sub.2, a Fd, an Fv fragment,
a disulfide stabilized Fv fragment (dsFv), a (dsFv).sub.2, a
bispecific dsFv (dsFv-dsFv'), a disulfide stabilized diabody (ds
diabody), a single-chain antibody molecule (scFv), an scFv dimer
(bivalent diabody), a bispecific scFv dimer, a multispecific
antibody, a heavy chain antibody, a camelized single domain
antibody, a nanobody, a domain antibody, or a bivalent domain
antibody. [1073] 45. The antibody or antigen binding fragment of
any of preceding embodiments, which is bispecific and comprises a
first antigen-binding domain and a second antigen-binding domain,
wherein the first and the second antigen-binding domains are
derived from any two monoclonal antibodies selected from the group
consisting of P2A-1A8, P2A-1A9, P2B-2G11, P2A-1A10, P2A-1B3,
P2B-2F6, P2B-2G4, P2C-1A3, P2C-1C8, P2C-1C10, P2C-1D5, P2C-1F11,
P2B-1G5, P2B-1A1, P2C-1D7, P2B-1A10, P2B-1D9, P2B-1E4, P2B-1G1,
P4A- 2D9, P5A-2G7, P5A-3C8, P5A-1D2, P5A-2F11, P5A-2E1, P2A-1A8,
P2A-1A9, P2B-2G11, P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4, P2C-1A3,
P2C-1C8, P2C-1C10, P2C-1D5, P2C- 1F11, P2B-1G5, P2B-1A1, P2C-1D7,
P2B-1A10, P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7, P5A-3C8,
P5A-1D2, P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6, P4B- 1F4,
P5A-1B6, P5A-1B8, P5A-1B9, P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9,
P5A-2H3, P5A-3A1, P5A-3A6, P5A-3B4, P5A-3C12, and P22A-1D1. [1074]
46. The antibody or antigen binding fragment of embodiment 45,
wherein the first and the second antigen-binding domains are
derived:
[1075] a) from P2C-1F11 and P2B-2F6, respectively;
[1076] b) from P2C-1F11 and P2A-1A8, respectively;
[1077] c) from P2C-1F11 and P2A-1A9, respectively;
[1078] d) from P2C-1F11 and P2B-2G11, respectively;
[1079] e) from P2C-1F11 and P2A-1A10, respectively;
[1080] f) from P2C-1F11 and P2A-1B3, respectively;
[1081] g) from P2C-1F11 and P2B-2G4, respectively;
[1082] h) from P2C-1F11 and P2C-1A3, respectively;
[1083] i) from P2C-1F11 and P2C-1C8, respectively;
[1084] j) from P2C-1F11 and P2C-1C10, respectively;
[1085] k) from P2C-1F11 and P2C-1D5, respectively;
[1086] l) from P2A-1A8 and P2A-1A9, respectively;
[1087] m) from P2A-1A8 and P2B-2G11, respectively;
[1088] n) from P2A-1A8 and P2A-1A10, respectively;
[1089] o) from P2A-1A8 and P2A-1B3, respectively;
[1090] p) from P2A-1A8 and P2B-2F6, respectively;
[1091] q) from P2A-1A8 and P2B-2G4, respectively;
[1092] r) from P2A-1A8 and P2C-1A3, respectively;
[1093] s) from P2A-1A8 and P2C-1C8, respectively;
[1094] t) from P2A-1A8 and P2C-1C10, respectively;
[1095] u) from P2A-1A8 and P2C-1D5, respectively;
[1096] v) from P2A-1A9 and 2B-2G11, respectively;
[1097] w) from P2A-1A9 and P2A-1A10, respectively;
[1098] x) from P2A-1A9 and P2A-1B3, respectively;
[1099] y) from P2A-1A9 and P2B-2F6, respectively;
[1100] z) from P2A-1A9 and P2B-2G4, respectively;
[1101] aa) from P2A-1A9 and P2C-1A3, respectively;
[1102] bb) from P2A-1A9 and P2C-1C8, respectively;
[1103] cc) from P2A-1A9 and P2C-1C10, respectively;
[1104] dd) from P2A-1A9 and P2C-1D5, respectively;
[1105] ee) from P2B-2G11 and P2A-1A10, respectively;
[1106] ff) from P2B-2G11 and P2A-1B3, respectively;
[1107] gg) from P2B-2G11 and P2B-2F6, respectively;
[1108] hh) from P2B-2G11 and P2B-2G4, respectively;
[1109] ii) from P2B-2G11 and P2C-1A3, respectively;
[1110] jj) from P2B-2G11 and P2C-1C8, respectively;
[1111] kk) from P2B-2G11 and P2C-1C10, respectively;
[1112] ll) from P2B-2G11 and P2C-1D5, respectively;
[1113] mm) from P2A-1A10 and P2A-1B3, respectively;
[1114] nn) from P2A-1A10 and P2B-2F6, respectively;
[1115] oo) from P2A-1A10 and P2B-2G4, respectively;
[1116] pp) from P2A-1A10 and P2C-1A3, respectively;
[1117] qq) from P2A-1A10 and P2C-1C8, respectively;
[1118] rr) from P2A-1A10 and P2C-1C10, respectively;
[1119] ss) from P2A-1A10 and P2C-1D5, respectively;
[1120] tt) from P2A-1B3 and P2B-2F6, respectively;
[1121] uu) from P2A-1B3 and P2B-2G4, respectively;
[1122] vv) from P2A-1B3 and P2C-1A3, respectively;
[1123] ww) from P2A-1B3 and P2C-1C8, respectively;
[1124] xx) from P2A-1B3 and P2C-1C10, respectively;
[1125] yy) from P2A-1B3 and P2C-1D5, respectively;
[1126] zz) from P2B-2F6 and P2B-2G4, respectively;
[1127] aaa) from P2B-2F6 and P2C-1A3, respectively;
[1128] bbb) from P2B-2F6 and P2C-1C8, respectively;
[1129] ccc) from P2B-2F6 and P2C-1C10, respectively;
[1130] ddd) from P2B-2F6 and P2C-1D5, respectively;
[1131] eee) from P2B-2G4 and P2C-1A3, respectively;
[1132] fff) from P2B-2G4 and P2C-1C8, respectively;
[1133] ggg) from P2B-2G4 and P2C-1C10, respectively;
[1134] hhh) from P2B-2G4 and P2C-1D5, respectively;
[1135] iii) from P2C-1A3 and P2C-1C8, respectively;
[1136] jjj) from P2C-1A3 and P2C-1C10, respectively;
[1137] kkk) from P2C-1A3 and P2C-1D5, respectively;
[1138] lll) from P2C-1C8 and P2C-1C10, respectively;
[1139] mmm) from P2C-1C8 and P2C-1D5, respectively; or
[1140] nnn) from P2C-1C10 and P2C-1D5, respectively. [1141] 47. The
antibody or antigen binding fragment of embodiments 1 or 2, which
is bispecific and comprises a first antigen-binding domain and a
second antigen-binding domain, wherein the first and the second
antigen-binding domains are derived from any two monoclonal
antibodies selected from the group consisting of P2C-1F11, P2B-2F6,
P2B-1G5, P2B-1A1, P2C-1D7, P2B-1A10, P2B-1D9, P2B-1E4, P2B-1G1,
P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2, P5A-2F11, P5A-2E1, P2A-1A8,
P2A-1A9, P2B-2G11, P2A-1A10, P2A-1B3, P2B-2F6, P2B-2G4, P2C-1A3,
P2C-1C8, P2C-1C10, P2C-1D5, P2C-1F11, P2B-1G5, P2B-1A1, P2C- 1D7,
P2B-1A10, P2B-1D9, P2B-1E4, P2B-1G1, P4A-2D9, P5A-2G7, P5A-3C8,
P5A-1D2, P5A-2F11, P5A-2E1, P5A-1C8, P1A-1C10, P4A-1H6, P4B-1F4,
P5A-1B6, P5A-1B8, P5A- 1B9, P5A-1D1, P5A-1D10, P5A-2D11, P5A-2G9,
P5A-2H3, P5A-3A1, P5A-3A6, P5A-3B4, P5A-3C12, and P22A-1D1. [1142]
48. The antibody or antigen binding fragment of embodiment 47,
wherein the first and the second antigen-binding domains are
derived:
[1143] a) from P2C-1F11 and P2B-1G5, respectively;
[1144] b) from P2C-1F11 and P2B-1A1, respectively;
[1145] c) from P2C-1F11 and P2C-1D7, respectively;
[1146] d) from P2C-1F11 and P2B-1A10, respectively;
[1147] e) from P2C-1F11 and P2B-1D9, respectively;
[1148] f) from P2C-1F11 and P2B-1E4, respectively;
[1149] g) from P2C-1F11 and P2B-1G1, respectively;
[1150] h) from P2C-1F11 and P4A-2D9, respectively;
[1151] i) from P2C-1F11 and P5A-2G7, respectively;
[1152] j) from P2C-1F11 and P5A-3C8, respectively;
[1153] k) from P2C-1F11 and P5A-1D2, respectively;
[1154] l) from P2C-1F11 and P5A-2F11, respectively;
[1155] m) from P2C-1F11 and P5A-2E1, respectively;
[1156] n) from P2C-1F11 and P5A-1C8, respectively;
[1157] o) from P2B-2F6 and P2B-1G5, respectively;
[1158] p) from P2B-2F6 and P2B-1A1, respectively;
[1159] q) from P2B-2F6 and P2C-1D7, respectively;
[1160] r) from P2B-2F6 and P2B-1A10, respectively;
[1161] s) from P2B-2F6 and P2B-1D9, respectively;
[1162] t) from P2B-2F6 and P2B-1E4, respectively;
[1163] u) from P2B-2F6 and P2B-1G1, respectively;
[1164] v) from P2B-2F6 and P4A-2D9, respectively;
[1165] w) from P2B-2F6 and P5A-2G7, respectively;
[1166] x) from P2B-2F6 and P5A-3C8, respectively;
[1167] y) from P2B-2F6 and P5A-1D2, respectively;
[1168] z) from P2B-2F6 and P5A-2F11, respectively;
[1169] aa) from P2B-2F6 and P5A-2E1, respectively;
[1170] bb) from P2B-2F6 and P5A-1C8, respectively;
[1171] cc) from P2B-1G5 and P2B-1A1, respectively;
[1172] dd) from P2B-1G5 and P2C-1D7, respectively;
[1173] ee) from P2B-1G5 and P2B-1A10, respectively;
[1174] ff) from P2B-1G5 and P2B-1D9, respectively;
[1175] gg) from P2B-1G5 and P2B-1E4, respectively;
[1176] hh) from P2B-1G5 and P2B-1G1, respectively;
[1177] ii) from P2B-1G5 and P4A-2D9, respectively;
[1178] jj) from P2B-1G5 and P5A-2G7, respectively;
[1179] kk) from P2B-1G5 and P5A-3C8, respectively;
[1180] ll) from P2B-1G5 and P5A-1D2, respectively;
[1181] mm) from P2B-1G5 and P5A-2F11, respectively;
[1182] nn) from P2B-1G5 and P5A-2E1, respectively;
[1183] oo) from P2B-1G5 and P5A-1C8, respectively;
[1184] pp) from P2B-1A1 and P2C-1D7, respectively;
[1185] qq) from P2B-1A1 and P2B-1A10, respectively;
[1186] rr) from P2B-1A1 and P2B-1D9, respectively;
[1187] ss) from P2B-1A1 and P2B-1E4, respectively;
[1188] tt) from P2B-1A1 and P2B-1G1, respectively;
[1189] uu) from P2B-1A1 and P4A-2D9, respectively;
[1190] vv) from P2B-1A1 and P5A-2G7, respectively;
[1191] ww) from P2B-1A1 and P5A-3C8, respectively;
[1192] xx) from P2B-1A1 and P5A-1D2, respectively;
[1193] yy) from P2B-1A1 and P5A-2F11, respectively;
[1194] zz) from P2B-1A15 and P5A-2E1, respectively;
[1195] aaa) from P2B-1A1 and P5A-1C8, respectively;
[1196] bbb) from P2C-1D7 and P2B-1A10, respectively;
[1197] ccc) from P2C-1D7 and P2B-1D9, respectively;
[1198] ddd) from P2C-1D7 and P2B-1E4, respectively;
[1199] eee) from P2C-1D7 and P2B-1G1, respectively;
[1200] fff) from P2C-1D7 and P4A-2D9, respectively;
[1201] ggg) from P2C-1D7 and P5A-2G7, respectively;
[1202] hhh) from P2C-1D7 and P5A-3C8, respectively;
[1203] iii) from P2C-1D7 and P5A-1D2, respectively;
[1204] jjj) from P2C-1D7 and P5A-2F11, respectively;
[1205] kkk) from P2B-1A15 and P5A-2E1, respectively;
[1206] lll) from P2B-1A1 and P5A-1C8, respectively;
[1207] mmm) from P2B-1A10 and P2B-1D9, respectively;
[1208] nnn) from P2B-1A10 and P2B-1E4, respectively;
[1209] ooo) from P2B-1A10 and P2B-1G1, respectively;
[1210] ppp) from P2B-1A10 and P4A-2D9, respectively;
[1211] qqq) from P2B-1A10 and P5A-2G7, respectively;
[1212] rrr) from P2B-1A10 and P5A-3C8, respectively;
[1213] sss) from P2B-1A10 and P5A-1D2, respectively;
[1214] ttt) from P2B-1A10 and P5A-2F1 1, respectively;
[1215] uuu) from P2B-1A10 and P5A-2E1, respectively;
[1216] vvv) from P2B-1A10 and P5A-1C8, respectively;
[1217] www) from P2B-1D9 and P2B-1E4, respectively;
[1218] xxx) from P2B-1D9 and P2B-1G1, respectively;
[1219] yyy) from P2B-1D9 and P4A-2D9, respectively;
[1220] zzz) from P2B-1D9 and P5A-2G7, respectively;
[1221] aaaa) from P2B-1D9 and P5A-3C8, respectively;
[1222] bbbb) from P2B-1D9 and P5A-1D2, respectively;
[1223] cccc) from P2B-1D9 and P5A-2F1 1, respectively;
[1224] dddd) from P2B-1D9 and P5A-2E1, respectively;
[1225] eeee) from P2B-1D9 and P5A-1C8, respectively;
[1226] ffff) from P2B-1E4 and P2B-1G1, respectively;
[1227] gggg) from P2B-1E4 and P4A-2D9, respectively;
[1228] hhhh) from P2B-1E4 and P5A-2G7, respectively;
[1229] iiii) from P2B-1E4 and P5A-3C8, respectively;
[1230] jjjj) from P2B-1E4 and P5A-1D2, respectively;
[1231] kkkk) from P2B-1E4 and P5A-2F1 1, respectively;
[1232] llll) from P2B-1E4 and P5A-2E1, respectively;
[1233] mmmm) from P2B-1E4 and P5A-1C8, respectively;
[1234] nnnn) from P2B-1G1 and P4A-2D9, respectively;
[1235] oooo) from P2B-1G1 and P5A-2G7, respectively;
[1236] pppp) from P2B-1G1 and P5A-3C8, respectively;
[1237] qqqq) from P2B-1G1 and P5A-1D2, respectively;
[1238] rrrr) from P2B-1G1 and P5A-2F11, respectively;
[1239] ssss) from P2B-1G1 and P5A-2E1, respectively;
[1240] tttt) from P2B-1G1 and P5A-1C8, respectively;
[1241] uuuu) from P4A-2D9 and P5A-2G7, respectively;
[1242] vvvv) from P4A-2D9 and P5A-3C8, respectively;
[1243] wwww) from P4A-2D9 and P5A-1D2, respectively;
[1244] xxxx) from P4A-2D9 and P5A-2F11, respectively;
[1245] yyyy) from P4A-2D9 and P5A-2E1, respectively;
[1246] zzzz) from P4A-2D9 and P5A-1C8, respectively;
[1247] aaaaa) from P5A-2G7 and P5A-3C8, respectively;
[1248] bbbbb) from P5A-2G7 and P5A-1D2, respectively;
[1249] ccccc) from P5A-2G7 and P5A-2F11, respectively;
[1250] ddddd) from P5A-2G7 and P5A-2E1, respectively;
[1251] eeeee) from P5A-2G7 and P5A-1C8, respectively;
[1252] fffff) from P5A-3C8 and P5A-1D2, respectively;
[1253] ggggg) from P5A-3C8 and P5A-2F11, respectively;
[1254] hhhhh) from P5A-3C8 and P5A-2E1, respectively;
[1255] iiiii) from P5A-3C8 and P5A-1C8, respectively;
[1256] jjjjj) from P5A-1D2 and P5A-2F11, respectively;
[1257] kkkkk) from P5A-1D2 and P5A-2E1, respectively;
[1258] lllll) from P5A-1D2 and P5A-1C8, respectively;
[1259] mmmmm) from P5A-2F11 and P5A-2E1, respectively;
[1260] nnnnn) from P5A-2F11 and P5A-1C8, respectively;
[1261] ooooo) from P5A-2E1 and P5A-1C8, respectively; [1262] 49.
The antibody or antigen binding fragment of any of the preceding
embodiments, which is a full human antibody. [1263] 50. The
antibody or antigen binding fragment of any of the preceding
embodiments, linked to one or more conjugate moieties. [1264] 51.
The antibody or antigen binding fragment of embodiment 50, wherein
the conjugate moiety comprises a therapeutic agent, a radioactive
isotope, a detectable label, a pharmacokinetic modifying moiety, or
a purifying moiety, and optionally the conjugate moiety is
covalently attached either directly or via a linker. [1265] 52. An
antibody or an antigen-binding fragment thereof, which competes for
binding to RBD of spike protein of SARS-CoV-2 with the antibody or
an antigen-binding fragment thereof of any one of embodiments 1-44.
[1266] 53. An isolated polynucleotide encoding the antibody or
antigen binding fragment of any of the embodiments 1-52. [1267] 54.
The isolated polynucleotide of embodiment 53, comprising a
nucleotide sequence selected from a group consisting of: SEQ ID
NOs: 9-10, 19-20, 29-30, 39-40, 49-50, 59-60, 63-64, 73-74, 83-84,
93-94, 103-104, 113-114, 144-145, 154-155, 164-165, 174-175,
184-185, 194-195, 204-205, 214-215, 224-225, 234-235, 244-245,
254-255, 264-265, 274-275, 284-285, 294-295, 304-305, 314-315,
324-325, 334-335, 344-345, 354-355, 364-365, 374-375, 384-385,
394-395, 404-405, 414-415, 424-425, and 434-435, or a homologous
sequence thereof having at least 80% sequence identity. [1268] 55.
The isolated polynucleotide of embodiment 54, wherein the homologue
sequence encodes the same protein as encoded by any nucleotide
sequence selected from the group consisting of SEQ ID NOs: 9-10,
19-20, 29-30, 39-40, 49-50, 59-60, 63-64, 73-74, 83-84, 93-94,
103-104, 113-114, 144-145, 154-155, 164-165, 174-175, 184-185,
194-195, 204-205, 214-215, 224-225, 234-235, 244-245, 254-255,
264-265, 274-275, 284-285, 294-295, 304-305, 314-315, 324-325,
334-335, 344-345, 354-355, 364-365, 374-375, 384-385, 394-395,
404-405, 414-415, 424-425, and 434-435. [1269] 56. A vector
comprising the isolated polynucleotide of any one of embodiments
53-55, optionally the vector is an expression vector. [1270] 57. A
host cell comprising the vector of embodiment 56. [1271] 58. A
pharmaceutical composition comprising the antibody or antigen
binding fragment of any one of embodiments 1-52, and a
pharmaceutically acceptable carrier, or comprising the
polynucleotide of claim 53, and a pharmaceutically acceptable
carrier. [1272] 59. The pharmaceutical composition of embodiment
58, comprising a combination of two or more antibodies or antigen
binding fragments of any one of embodiments 1-52, and a
pharmaceutically acceptable carrier. [1273] 60. The pharmaceutical
composition of embodiment 59, wherein the two or more antibodies or
the antigen binding fragments thereof bind to different epitopes in
RBD of spike protein of SARS-CoV-2. [1274] 61. The pharmaceutical
composition of embodiment 60, wherein the two or more antibodies
comprise a first antibody and a second antibody selected from the
group consisting of P2A-1A8, P2A-1A9, P2B-2G11, P2A-1A10, P2A-1B3,
P2B-2F6, P2B-2G4, P2C-1A3, P2C-1C8, P2C-1C10, P2C-1D5, P2C-1F11,
P2B-1G5, P2B-1A1, P2C-1D7, P2B-1A10, P2B-1D9, P2B- 1E4, P2B-1G1,
P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2, P5A-2F11, P5A-2E1, P5A-1C8,
P1A-1C10, P4A-1H6, P4B-1F4, P5A-1B6, P5A-1B8, P5A-1B9, P5A-1D1,
P5A-1D10, P5A- 2D11, P5A-2G9, P5A-2H3, P5A-3A1, P5A-3A6, P5A-3B4,
P5A-3C12, and P22A-1D1, or an antigen binding fragment thereof.
[1275] 62. The pharmaceutical composition of embodiment 60, wherein
the two or more antibodies comprise a first antibody and a second
antibody selected from the group consisting of P2C-1F11, P2B-2F6,
P2B-1G5, P2B-1A1, P2C-1D7, P2B-1A10, P2B-1D9, P2B-1E4, P2B-1G1,
P4A-2D9, P5A-2G7, P5A-3C8, P5A-1D2, P5A-2F11, P5A-2E1, and P5A-1C8,
or an antigen binding fragment thereof. [1276] 63. The
pharmaceutical composition of embodiment 60, wherein the two or
more antibodies comprise a first antibody which comprises P2C-1F11
or an antigen binding fragment thereof, and a second antibody which
is selected from the group consisting of P2C-1A3, P2C-1C10,
P2B-2F6, P2B-1G5, and P2A-1B3, or an antigen binding fragment
thereof, optionally, the pharmaceutical composition comprises a
first antibody comprising heavy chain CDR sequences and light chain
CDR sequences derived from P2C-1F11, and a second antibody
comprising heavy chain CDR sequences and light chain CDR sequences
derived from antibody P2B-2F6. [1277] 64. The pharmaceutical
composition of embodiment 60, wherein the two or more antibodies
comprise a first antibody which comprises P2C-1A3 or an antigen
binding fragment thereof, and a second antibody which is selected
from the group consisting of P2C-1F11, and P2A-1B3, or an antigen
binding fragment thereof. [1278] 65. The pharmaceutical composition
of embodiment 60, wherein the two or more antibodies comprise a
first antibody which comprises P2B-2F6 or an antigen binding
fragment thereof, and a second antibody selected from the group
consisting of P2C-1C10, P2C-1F11, P2B-1G5, and P2A-1B3, or an
antigen binding fragment thereof. [1279] 66. The pharmaceutical
composition of embodiment 60, wherein the two or more antibodies
comprise a first antibody which comprises P2A-1B3 or an antigen
binding fragment thereof, and a second antibody selected from the
group consisting of P2C-1A3, P2C-1C10, P2C-1F11, P2B-2F6, and
P2A-1A10, or an antigen binding fragment thereof. [1280] 67. A
method of producing the antibody or antigen binding fragment of any
of embodiments 1-52 comprising culturing the host cell of
embodiment 57 under the condition at which the vector of embodiment
56 is expressed. [1281] 68. The method of embodiment 67, further
comprising purifying the antibody produced by the host cell. [1282]
69. A kit for detecting a SARS-CoV-2 antigen, comprising the
antibody or antigen binding fragment of any of embodiments 1-52.
[1283] 70. The kit of embodiment 69, further comprising a control
reagent comprising RBD of spike protein of the SARS-CoV-2,
optionally, the kit further comprises a set of reagents for
detecting complex of the antibody or the antigen-binding fragment
bound to the SARS-CoV-2 antigen. [1284] 71. A method of treating
SARS-CoV-2 infection or a disease, disorder or condition associated
with SARs-CoV-2 infection in a subject, comprising administering a
therapeutically effective amount of one or more of the antibody or
antigen binding fragment of any of embodiments 1-52, or of one or
more of the polynucleotides of any embodiments 53-55, or of one or
more of the vectors of embodiment 56, or of the pharmaceutical
composition of any of embodiments 58-66 to the subject. [1285] 72.
A method of preventing SARS-CoV-2 infection or a disease, disorder
or condition associated with SARs-CoV-2 infection in a subject,
comprising administering a therapeutically effective amount of one
or more of the antibody or antigen binding fragment of any of
embodiments 1-52, or of one or more of the polynucleotides of any
embodiments 53-55, or of one or more of the vectors of embodiment
56, or of the pharmaceutical composition of any of embodiments
58-66 to the subject. [1286] 73. The method of embodiments 71 or
72, wherein the administration is via oral, nasal, intravenous,
subcutaneous, or intramuscular administration. [1287] 74. The
method of embodiment 73, wherein the subject is human. [1288] 75.
The method of any of embodiments 71-74, further comprising
administering a therapeutically effective amount of a second
bioactive agent, optionally the second bioactive agent is a
therapeutic agent or a prophylactic agent. [1289] 76. The method of
embodiment 75, wherein the therapeutic agent is an anti-viral
agent, optionally, the anti-viral agent comprises an antiviral
peptide, an anti-viral antibody, an anti-viral compound, an
anti-viral cytokine, or an anti-viral oligonucleotide. [1290] 77. A
method of detecting presence or amount of SARS-CoV-2 virus antigen
in a sample, comprising contacting the sample with one or more of
the antibody or antigen binding fragment of any of embodiments
1-52, and determining the presence or the amount of the SARS-CoV-2
virus antigen in the sample. [1291] 78. Use of one or more of the
antibody or antigen binding fragment of any of embodiments 1-52 in
the manufacture of a medicament for treating SARS-CoV-2 infection
or a disease, disorder or condition associated with SARs-CoV-2
infection. [1292] 79. Use of one or more of the antibody or antigen
binding fragment of any of embodiments 1-52 in the manufacture of a
diagnostic reagent for detecting SARS-CoV-2 infection. [1293] 80. A
kit for detecting an antibody capable of specifically binding to
receptor-binding domain (RBD) of the spike protein of SARS-CoV-2,
comprising a polypeptide comprising an amino acid sequence
comprising SEQ ID NO: 128. [1294] 81. The kit of embodiment 80,
wherein the polypeptide is immobilized on a substrate. [1295] 82.
The kit of embodiments 81, further comprising a set of reagents for
detecting complex of the antibody bound to the polypeptide. [1296]
83. A method of detecting presence or amount of an antibody capable
of specifically binding to RBD of the spike protein of SARS-CoV-2
in a sample, comprising contacting the sample with a polypeptide
comprising an amino acid sequence comprising SEQ ID NO: 128, and
determining the presence or the level of the antibody in the
sample. [1297] 84. The method of embodiment 83, wherein the absence
of the antibody in the sample or the level of the antibody in the
sample being below a threshold indicates that the subject is more
likely to suffer from disease progression. [1298] 85. A method of
determining the likelihood of disease progression in a subject
infected with SARS-CoV-2, the method comprising: contacting a
sample obtained from the subject with a polypeptide comprising an
amino acid sequence comprising SEQ ID NO: 128, and detecting the
presence or the level of an antibody in the sample wherein the
antibody is capable of specifically binding to RBD of the spike
protein of the SARS-CoV-2, wherein the subject is likely to
experience disease progression when the antibody in the sample is
absent or is below a threshold. [1299] 86. A method of monitoring
treatment response in a subject infected with SARS-CoV-2 and
received a treatment, the method comprising:
[1300] (i) contacting a sample from the subject with a peptide
comprising an amino acid sequence comprising SEQ ID NO: 128;
[1301] (ii) detecting a first level of an antibody in the sample
wherein the antibody is capable of specifically binding to RBD of
the spike protein of the SARS-CoV-2; and
[1302] (iii) comparing the first level of the antibody with a
second level of the antibody detected in the subject prior to the
treatment;
[1303] wherein the first level being higher than the second level
indicates that the subject is responsive to the treatment. [1304]
87. A method of neutralizing SARS-CoV-2 in a subject or in a sample
in vitro, comprising administering a therapeutically effective
amount of one or more of the antibody or antigen binding fragment
of any of embodiments 1-52, or the pharmaceutical composition of
any of claims 58-66 to the subject or to the sample. [1305] 88. A
crystal of RBD of the spike protein of SARS-CoV-2 in complex with
an antibody. [1306] 89. The crystal of embodiment 88, having or
consisting of a P2.sub.12.sub.12.sub.1 space group with unit cell
dimensions of a=70.23 .ANG., b=90.15 .ANG., and c=112.35 .ANG.,
having or consisting of a C121 space group with unit cell
dimensions of a=194.88 .ANG., b=85.39 .ANG., and c=58.51 .ANG.,
having or consisting of a C2 space group with unit cell dimensions
of a=193.34 .ANG., b=86.60 .ANG., and c=57.16 .ANG., having or
consisting of a C2 space group with unit cell dimensions of
a=158.75 .ANG., b=67.51 .ANG., and c=154.37 .ANG., or having or
consisting of a P2.sub.12.sub.12.sub.1 space group with unit cell
dimensions of a=112.54 .ANG., b=171.57 .ANG., and c=54.87 .ANG..
[1307] 90. The crystal of embodiment 88, wherein the antibody
comprises a heavy chain variable region of SEQ ID NO: 47 and a
light chain variable region of SEQ ID NO: 48. [1308] 91. The
crystal of embodiment 88, wherein the antibody comprises a heavy
chain variable region of SEQ ID NO: 111 and a light chain variable
region of SEQ ID NO: 112. [1309] 92. The crystal of embodiments 88,
wherein the antibody comprises a heavy chain variable region of SEQ
ID NO: 432 and a light chain variable region of SEQ ID NO: 433.
[1310] 93. The crystal of embodiments 88, wherein the antibody
comprises a heavy chain variable region of SEQ ID NO: 242 and a
light chain variable region of SEQ ID NO: 243. [1311] 94. The
crystal of embodiments 88, wherein the antibody comprises a heavy
chain variable region of SEQ ID NO: 232 and a light chain variable
region of SEQ ID NO: 233. [1312] 95. A computer-implemented method
for causing a display of a graphical three-dimensional
representation of the structure of a portion of a crystal of RBD of
the spike protein of SARS-CoV-2 in complex with an anti-SARS-CoV-2
antibody or an antigen-binding fragment thereof, wherein the method
comprises:
[1313] causing said display of said graphical three-dimensional
representation by a computer system programmed with instructions
for transforming structure coordinates into said graphical
three-dimensional representation of said structure and for
displaying said graphical three-dimensional representation,
[1314] wherein said graphical three-dimensional representation is
generated by transforming said structure coordinates into said
graphical three-dimensional representation of said structure,
[1315] wherein said structure coordinates comprise structure
coordinates of the backbone atoms of the portion of the
crystal,
[1316] wherein the portion of the crystal comprises a RBD binding
site, and
[1317] wherein the crystal has the space group symmetry
P2.sub.12.sub.12.sub.1 or C121. [1318] 96. The computer-implemented
method of embodiment 95, wherein the RBD comprises an amino acid
sequence as shown in SEQ ID NO: 124, and the antibody comprises: a)
a heavy chain variable region of SEQ ID NO: 47 and a light chain
variable region of SEQ ID NO: 48; or b) a heavy chain variable
region of SEQ ID NO: 111 and a light chain variable region of SEQ
ID NO: 112; or c) a heavy chain variable region of SEQ ID NO: 432
and a light chain variable region of SEQ ID NO: 433; or d) a heavy
chain variable region of SEQ ID NO: 242 and a light chain variable
region of SEQ ID NO: 243; or e) a heavy chain variable region of
SEQ ID NO: 232 and a light chain variable region of SEQ ID NO: 233.
[1319] 97. The computer-implemented method of embodiment 95,
wherein the structure coordinates comprise the structure
coordinates of the backbone atoms of the amino acid residues
corresponding to K444, G446, G447, N448, Y449, N450, L452, V483,
E484, G485, F490 and/or S494 of the RBD, wherein the residue
numbering is according to SEQ ID NO: 134. [1320] 98. The
computer-implemented method of embodiment 95, wherein the structure
coordinates comprise the structure coordinates of the backbone
atoms of the amino acid residues corresponding to Y453, L455, F456,
R457, K458, 5459, N460, Y473, A475, G476, S477, F486, N487, Y489,
Q493, G502, Y505, R403, T415, G416, K417, D420 and/or Y421 of the
RBD, wherein the residue numbering is according to SEQ ID NO: 134.
[1321] 99. The computer-implemented method of embodiment 95,
wherein the structure coordinates comprise the structure
coordinates of the backbone atoms of the amino acid residues
corresponding to T415, G416, K417, D420, Y421, L455, F456, R457,
K458, N460, Y473, A475, G476, S477, F486, N487, Y489 and/or Q493 of
the RBD, wherein the residue numbering is according to SEQ ID NO:
134. [1322] 100. The computer-implemented method of embodiment 95,
wherein the structure coordinates comprise the structure
coordinates of the backbone atoms of the amino acid residues
corresponding to T415, G416, K417, D420, Y421, Y453, L455, F456,
R457, K458, N460, Y473, Q474, A475, G476, S477, N487, Y489, Q493
and/or Y505 of the RBD, wherein the residue numbering is according
to SEQ ID NO: 134. [1323] 101. The computer-implemented method of
embodiment 95, wherein the structure coordinates comprise the
structure coordinates of the backbone atoms of the amino acid
residues corresponding to T415, G416, K417, D420, Y421, Y453, L455,
F456, R457, K458, N460, Y473, A475, G476, S477, F486, N487, Y489
and/or Q493 of the RBD, wherein the residue numbering is according
to SEQ ID NO: 134. [1324] 102. The computer-implemented method of
embodiment 95, wherein the structure coordinates comprise the
structure coordinates of the backbone atoms of the amino acid
residues corresponding to T415, G416, K417, D420, Y421, L455, F456,
R457, K458, N460, Y473, Q474, A475, G476, S477, F486, N487, Y489
and/or Q493 of the RBD, wherein the residue numbering is according
to SEQ ID NO: 134. [1325] 103. A machine-readable data storage
medium comprising a data storage material encoded with
machine-readable instructions for:
[1326] (a) transforming data into a graphical three-dimensional
representation for the structure of a portion of a crystal of RBD
of the spike protein of SARS-CoV-2 in complex with an
anti-SARS-CoV-2 antibody or an antigen-binding fragment thereof;
and
[1327] (b) causing the display of said graphical three-dimensional
representation;
[1328] wherein said data comprise structure coordinates of the
backbone atoms of the amino acids defining a RBD binding site; and
wherein the crystal or structural homolog has the space group
symmetry P2.sub.12.sub.12.sub.1 or C121. [1329] 104. A method of
screening for molecules that may be a binding molecule of RBD of
the spike protein of SARS-CoV-2, comprising:
[1330] (a) computationally screening agents against a
three-dimensional model to identify potential binding molecules of
the RBD;
[1331] wherein the three-dimensional model comprises a
three-dimensional model of at least a portion of a crystal of RBD
of the spike protein of SARS-CoV-2 in complex with an
anti-SARS-CoV-2 antibody or an antigen-binding fragment
thereof;
[1332] wherein the three dimensional model is generated from at
least a portion of the structure coordinates of the crystal by a
computer algorithm for generating a three-dimensional model of the
crystal useful for identifying agents that are potential binding
molecules of the RBD; wherein the crystal comprises a polypeptide
comprising an amino acid sequence SEQ ID NO: 124, and further
comprises an antibody comprising: a) a heavy chain variable region
of SEQ ID NO: 47 and a light chain variable region of SEQ ID NO:
48, orb) a heavy chain variable region of SEQ ID NO: 111 and a
light chain variable region of SEQ ID NO: 112; and
[1333] wherein the crystal diffracts x-rays for the determination
of atomic coordinates to a resolution of 5 .ANG. or better. [1334]
105. A method for obtaining structural information about a molecule
or molecular complex comprising applying at least a portion of the
structure coordinates of a RBD of the spike protein of SARS-CoV-2
in complex with an anti-SARS-CoV-2 antibody or an antigen-binding
fragment thereof, to an X-ray diffraction pattern of the molecule
or molecular complex's crystal structure to cause the generation of
a three-dimensional electron density map of at least a portion of
the molecule or molecular complex;
[1335] wherein the crystal comprises a polypeptide comprising an
amino acid sequence SEQ ID NO: 124, and further comprises an
antibody comprising: a) a heavy chain variable region of SEQ ID NO:
47 and a light chain variable region of SEQ ID NO: 48, orb) a heavy
chain variable region of SEQ ID NO: 111 and a light chain variable
region of SEQ ID NO: 112,
[1336] wherein the crystal diffracts x-rays for the determination
of atomic coordinates to a resolution of 5 .ANG. or better. [1337]
106. Use of a composition comprising a modified antibody or an
antigen-binding fragment thereof and one or more pharmaceutically
acceptable carriers for manufacturing a medicament for treating or
preventing a disease, wherein the composition comprises said
modified antibody or said antigen-binding fragment thereof that
comprises at least an antigen-binding domain having an
antigen-binding affinity and a covalently linked modified human IgG
constant domain, wherein said antigen-binding affinity comprises
SARS-CoV-2 binding affinity, said antigen-binding affinity
comprises at least 50% less or non-detectable binding affinity to
SARS-CoV or MERS-CoV compared to said SARS-CoV-2 binding affinity,
and wherein said modified human IgG constant domain comprises a
substitution with tyrosine at amino acid residue 252, a
substitution with threonine at amino acid residue 254, and a
substitution with glutamic acid at amino acid residue 256, numbered
according to the EU index as in Kabat, said modified antibody has
an increased affinity for FcRn compared to the affinity to FcRn of
an antibody having a wild type human IgG constant domain, and
wherein said disease is caused by said SARS-CoV-2 or related to
infection of said SARS-CoV-2 in said subject. [1338] 107. The use
of embodiment 106, wherein said subject is a symptomatic
non-hospitalized adult with COVID-19 caused by SARS-CoV-2
infection, is aged 60 years and older, is any age having at least
one of the following conditions selected from smoking, has
exogenous or endogenous immunosuppression having HIV infection with
CD4 count <200 cells/mm.sup.3, receives corticosteroids
equivalent to prednisone .gtoreq.20 mg daily for at least 14
consecutive days within 30 days prior to the treatment, has a
treatment with one or more biologics therapeutical agents, one or
more immunomodulators, cancer chemotherapy within 90 days prior to
the treatment; has chronic lung disease, chronic asthma; obesity
with body mass index [BMI]>35, symptoms of COVID-19 selected
from fever, cough, sore throat, malaise, headache, muscle pain,
nausea, vomiting, diarrhea, loss of taste and smell, or a
combination thereof, has shortness of breath, dyspnea, or abnormal
chest imaging, having evidence of lower respiratory disease during
clinical assessment or imaging, has saturation of oxygen (SpO2)
.gtoreq.94% on room air at sea level, has severe symptoms of the
infection of said SARS-CoV-2, having SpO2 <94% on room air at
sea level, having a ratio of arterial partial pressure of oxygen to
fraction of inspired oxygen (PaO2/FiO2) <300 mmHg, respiratory
frequency>30 breaths per minute, lung infiltrates >50%,
active symptoms of antibody-dependent enhancement (ADE), a history
of antibody-dependent enhancement (ADE), being allergic to an
antibody treatment, being a hospital inpatient requiring supportive
management of complications of severe infection of said SARS-CoV-2
selected from pneumonia, has hypoxemic respiratory failure/ARDS,
sepsis and septic shock, cardiomyopathy and arrhythmia, acute
kidney injury, and complications from prolonged hospitalization,
including secondary bacterial and fungal infections,
thromboembolism, gastrointestinal bleeding, critical illness
polyneuropathy/myopathy, or a combination thereof.
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Sequence CWU 1
1
43718PRTHomo sapiensP2A-1A8 HCDR1(1)..(8) 1Gly Phe Ala Phe Asp Asp
Tyr Ala1 528PRTHomo sapiensP2A-1A8 HCDR2(1)..(8) 2Ser Thr Trp Asn
Ser Gly Thr Ile1 5323PRTHomo sapiensP2A-1A8 HCDR3(1)..(23) 3Ala Lys
Leu Gly Gly Tyr Ser Asp Tyr Asp Tyr Pro Arg Pro Gly Asp1 5 10 15His
Tyr Tyr Gly Leu Asp Val 2049PRTHomo sapiensP2A-1A8 LCDR1(1)..(9)
4Ser Ser Asp Val Gly Ser Tyr Asn Leu1 554PRTHomo sapiensP2A-1A8
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 5Asp Val Asn
Xaa1610PRTHomo sapiensP2A-1A8 LCDR3(1)..(10) 6Arg Ser Tyr Thr Asp
Ser Asn Thr Tyr Val1 5 107130PRTHomo sapiensP2A-1A8 VH(1)..(130)
7Glu Val Gln Leu Val Glu Ser Gly Gly Asp Leu Val Gln Pro Gly Arg1 5
10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ala Phe Asp Asp
Tyr 20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu
Trp Val 35 40 45Ser Gly Ser Thr Trp Asn Ser Gly Thr Ile Ala Tyr Ala
Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys
Lys Ser Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Thr Glu Asp
Thr Ala Leu Tyr Tyr Cys 85 90 95Ala Lys Leu Gly Gly Tyr Ser Asp Tyr
Asp Tyr Pro Arg Pro Gly Asp 100 105 110His Tyr Tyr Gly Leu Asp Val
Trp Gly Gln Gly Thr Thr Val Thr Val 115 120 125Ser Ser
1308110PRTHomo sapiensP2A-1A8 VL(1)..(110) 8Gln Ser Ala Leu Thr Gln
Pro Ala Ser Val Ser Gly Ser Pro Gly Gln1 5 10 15Ser Ile Thr Ile Ser
Cys Thr Gly Thr Ser Ser Asp Val Gly Ser Tyr 20 25 30Asn Leu Val Ser
Trp Tyr Gln Gln His Pro Gly Lys Val Pro Lys Leu 35 40 45Leu Ile Tyr
Asp Val Asn Lys Arg Pro Ser Gly Ile Ser Asn Arg Phe 50 55 60Ser Gly
Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75
80Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Arg Ser Tyr Thr Asp Ser
85 90 95Asn Thr Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu 100
105 1109390DNAHomo sapiensP2A-1A8 VHnu(1)..(390) 9gaagtgcagc
tggtggagtc tgggggagac ttggtacagc ctggcaggtc cctgagactc 60tcctgcgcag
cctctggatt cgcctttgat gattatgcca tgcactgggt ccggcaagct
120ccagggaagg gcctggagtg ggtctcaggt agtacttgga atagtgggac
catagcctat 180gcggactctg tgaagggccg attcaccatc tccagagaca
acgccaagaa gtccctgtat 240ctgcaaatga acagtctgag aactgaggac
acggccttat attactgtgc aaagttgggg 300ggctacagtg actacgatta
cccgaggccg ggagaccact attacggttt ggacgtctgg 360ggccaaggga
ccacggtcac cgtctcctca 39010330DNAHomo sapiensP2A-1A8 VLnu(1)..(330)
10cagtctgccc tgactcagcc tgcctccgtg tctgggtctc ctggacagtc gatcaccatc
60tcctgcactg gaaccagcag tgatgttggg agttataacc ttgtctcctg gtaccaacag
120cacccaggca aagtccccaa actcttgatt tatgatgtca ataagcggcc
ctcagggatt 180tccaatcgct tctctggctc caagtctggc aacacggcct
ccctgaccat ctctgggctc 240caggctgagg acgaggctga ttattactgc
agatcatata cagacagcaa cacttatgtc 300ttcggaactg ggaccaaggt
caccgtccta 330118PRTHomo sapiensP2A-1A9/P2B-2G11 HCDR1(1)..(8)
11Gly Phe Thr Phe Asp Asp Tyr Ala1 5128PRTHomo
sapiensP2A-1A9/P2B-2G11 HCDR2(1)..(8) 12Ile Ser Trp Asn Gly Gly Ile
Ile1 51317PRTHomo sapiensP2A-1A9/P2B-2G11 HCDR3(1)..(17) 13Ala Lys
Val Ala Gly Arg Gly Asp Tyr Asp Tyr Tyr Tyr Gly Met Asp1 5 10
15Val149PRTHomo sapiensP2A-1A9/P2B-2G11 LCDR1(1)..(9) 14Ser Ser Asn
Ile Gly Ala Gly Tyr Asp1 5154PRTHomo sapiensP2A-1A9/P2B-2G11
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 15Gly Asn Asn
Xaa11611PRTHomo sapiensP2A-1A9/P2B-2G11 LCDR3(1)..(11) 16Gln Ser
Tyr Asp Ser Ser Leu Ser Gly Ser Val1 5 1017124PRTHomo
sapiensP2A-1A9 VH(1)..(124) 17Glu Val Gln Leu Val Glu Ser Gly Gly
Gly Leu Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30Ala Met His Trp Val Arg Gln
Val Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Gly Ile Ser Trp Asn
Gly Gly Ile Ile Gly Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ala Lys Thr Ser Leu Tyr65 70 75 80Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Ala Lys
Val Ala Gly Arg Gly Asp Tyr Asp Tyr Tyr Tyr Gly Met Asp 100 105
110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115
12018111PRTHomo sapiensP2A-1A9 VL(1)..(111) 18Gln Ser Val Leu Thr
Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln1 5 10 15Arg Val Thr Ile
Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly 20 25 30Tyr Asp Val
His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45Leu Ile
Tyr Gly Asn Asn Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu65 70 75
80Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser
85 90 95Leu Ser Gly Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 11019372DNAHomo sapiensP2A-1A9 VHnu(1)..(372) 19gaagtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggcaggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttgat gattatgcca tgcactgggt ccggcaagtt
120ccagggaagg gcctggagtg ggtctcaggt attagttgga atggtggtat
cataggctac 180gcggactctg tgaagggccg attcaccatc tccagagaca
acgccaagac ttccctgtat 240ctgcaaatga acagtctgag agctgaggac
acggccttgt attactgtgc aaaagtcgcg 300ggaagggggg attacgacta
ttactatggt atggacgtct ggggccaagg gaccacggtc 360accgtctcct ca
37220333DNAHomo sapiensP2A-1A9 VLnu(1)..(333) 20cagtctgtgc
tgacgcagcc gccctcagtg tctggggccc cagggcagag ggtcaccatc 60tcctgcactg
ggagcagctc caacatcggg gcaggttatg atgtacactg gtaccagcaa
120cttccaggaa cagcccccaa actcctcatc tatggtaaca acaatcgccc
ctcaggggtc 180cctgaccgat tctctggctc caagtctggc acctcagcct
ccctggccat cactgggctc 240caggctgagg atgaggctga ttattactgc
cagtcctatg acagcagcct gagtggttcg 300gtattcggcg gagggaccaa
gctgaccgtc cta 333218PRTHomo sapiensP2A-1A10 HCDR1(1)..(8) 21Gly
Tyr Thr Phe Thr Gly Tyr Tyr1 5228PRTHomo sapiensP2A-1A10
HCDR2(1)..(8) 22Ile Asn Pro Asn Ser Gly Gly Thr1 52319PRTHomo
sapiensP2A-1A10 HCDR3(1)..(19) 23Ala Arg Val Pro Tyr Cys Ser Ser
Thr Ser Cys His Arg Asp Trp Tyr1 5 10 15Phe Asp Leu2412PRTHomo
sapiensP2A-1A10 LCDR1(1)..(12) 24Gln Ser Leu Leu Asp Ser Asp Asp
Gly Asn Thr Tyr1 5 10254PRTHomo sapiensP2A-1A10
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 25Thr Leu Ser
Xaa1269PRTHomo sapiensP2A-1A10 LCDR3(1)..(9) 26Met Gln Arg Ile Glu
Phe Pro Leu Thr1 527126PRTHomo sapiensP2A-1A10 VH(1)..(126) 27Gln
Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1 5 10
15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr
20 25 30Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp
Met 35 40 45Gly Arg Ile Asn Pro Asn Ser Gly Gly Thr Asn Tyr Ala Gln
Lys Phe 50 55 60Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser
Thr Ala Tyr65 70 75 80Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr
Ala Val Tyr Tyr Cys 85 90 95Ala Arg Val Pro Tyr Cys Ser Ser Thr Ser
Cys His Arg Asp Trp Tyr 100 105 110Phe Asp Leu Trp Gly Arg Gly Thr
Leu Val Thr Val Ser Ser 115 120 12528113PRTHomo sapiensP2A-1A10
VL(1)..(113) 28Asp Ile Val Met Thr Gln Thr Pro Leu Ser Leu Pro Val
Thr Pro Gly1 5 10 15Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser
Leu Leu Asp Ser 20 25 30Asp Asp Gly Asn Thr Tyr Leu Asp Trp Tyr Leu
Gln Lys Pro Gly Gln 35 40 45Ser Pro Gln Leu Leu Ile Tyr Thr Leu Ser
Tyr Arg Ala Ser Gly Val 50 55 60Pro Asp Arg Phe Ser Gly Ser Gly Ser
Gly Thr Asp Phe Thr Leu Lys65 70 75 80Ile Ser Arg Val Glu Ala Glu
Asp Val Gly Val Tyr Tyr Cys Met Gln 85 90 95Arg Ile Glu Phe Pro Leu
Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 100 105 110Lys29378DNAHomo
sapiensP2A-1A10 VHnu(1)..(378) 29caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtc 60tcctgcaagg cttctggata caccttcacc
ggctactata tgcactgggt gcgacaggcc 120cctggacaag ggcttgagtg
gatgggacgg atcaacccta acagtggtgg cacaaactat 180gcacagaagt
ttcagggcag ggtcaccatg accagggaca cgtccatcag cacagcctac
240atggagctga gcaggctgag atctgacgac acggccgtgt attactgtgc
gagagtcccc 300tattgtagta gtaccagctg ccatcgggac tggtacttcg
atctctgggg ccgtggcacc 360ctggtcactg tctcctca 37830339DNAHomo
sapiensP2A-1A10 VLnu(1)..(339) 30gatattgtga tgacccagac tccactctcc
ctgcccgtca cccctggaga gccggcctcc 60atctcctgca ggtctagtca gagcctcttg
gatagtgatg atggaaacac ctatttggac 120tggtacctgc agaagccagg
gcagtctcca cagctcctga tctatacgct ttcctatcgg 180gcctctggag
tcccagacag gttcagtggc agtgggtcag gcactgattt cacactgaaa
240atcagcaggg tggaggctga ggatgttgga gtttattact gcatgcaacg
tatagagttt 300ccgctcactt tcggcggagg gaccaaggtg gagatcaaa
339318PRTHomo sapiensP2A-1B3 HCDR1(1)..(8) 31Gly Phe Ser Phe Asn
Arg Tyr Ser1 5328PRTHomo sapiensP2A-1B3 HCDR2(1)..(8) 32Ile Ser Ala
Ser Gly Asn Thr Ile1 53316PRTHomo sapiensP2A-1B3 HCDR3(1)..(16)
33Ala Arg Pro Ala Met Val Arg Glu Gly Thr Tyr Asn Trp Phe Asp Pro1
5 10 15347PRTHomo sapiensP2A-1B3 LCDR1(1)..(7) 34Gln Ser Val Ser
Asn Asp Tyr1 5354PRTHomo sapiensP2A-1B3
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 35Tyr Ala Ser
Xaa13610PRTHomo sapiensP2A-1B3 LCDR3(1)..(10) 36Gln Gln Tyr Gly Asp
Ser Pro Pro Ile Thr1 5 1037123PRTHomo sapiensP2A-1B3 VH(1)..(123)
37Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly1
5 10 15Ser Leu Arg Leu Ser Cys Val Ala Ser Gly Phe Ser Phe Asn Arg
Tyr 20 25 30Ser Met Asn Trp Leu Arg Gln Thr Pro Arg Lys Gly Leu Glu
Trp Leu 35 40 45Ser Tyr Ile Ser Ala Ser Gly Asn Thr Ile Tyr Tyr Ala
Asp Ser Val 50 55 60Arg Gly Arg Phe Thr Thr Ser Arg Asp Asn Ala Lys
Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Asp Asp Asp
Thr Ala Val Tyr Phe Cys 85 90 95Ala Arg Pro Ala Met Val Arg Glu Gly
Thr Tyr Asn Trp Phe Asp Pro 100 105 110Trp Gly Gln Gly Thr Leu Val
Thr Val Ser Ser 115 12038109PRTHomo sapiensP2A-1B3 VL(1)..(109)
38Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1
5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Asn
Asp 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg
Leu Leu 35 40 45Ile Tyr Tyr Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp
Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
Ser Arg Leu Glu65 70 75 80Pro Gly Asp Ser Ala Val Tyr Tyr Cys Gln
Gln Tyr Gly Asp Ser Pro 85 90 95Pro Ile Thr Phe Gly Gln Gly Thr Arg
Leu Glu Ile Lys 100 10539369DNAHomo sapiensP2A-1B3 VHnu(1)..(369)
39gaggtgcagc tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctcagactc
60tcctgtgtcg cctctggatt ctccttcaat cgatatagta tgaattggct ccgccagact
120ccacggaagg ggctggagtg gctttcatac atcagtgcca gtggaaacac
catatactac 180gctgactctg tgaggggccg attcaccacc tccagagaca
atgccaagaa cacactgtat 240ctgcaaatga acagcctgcg agacgacgac
acggctgtct atttctgtgc gcgacccgct 300atggttcggg aggggaccta
caactggttc gacccctggg gccagggaac cctggtcacc 360gtctcctca
36940327DNAHomo sapiensP2A-1B3 VLnu(1)..(327) 40gaaattgtgt
tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60ctctcctgca
gggccagtca gagtgttagc aacgactact tagcctggta ccagcagaaa
120cctggccagg ctcccaggct cctcatctac tatgcatcca gcagggccac
tggcatccca 180gacaggttca gtggcagtgg gtctgggaca gacttcactc
tcaccatcag cagactggag 240cctggagatt ctgcagtgta ttactgtcag
cagtatggtg actcacctcc gatcaccttc 300ggccaaggga cacgactgga gattaaa
327419PRTHomo sapiensP2B-2F6 HCDR1(1)..(9) 41Gly Tyr Ser Ile Ser
Ser Gly Tyr Tyr1 5427PRTHomo sapiensP2B-2F6 HCDR2(1)..(7) 42Ile Tyr
His Ser Gly Ser Thr1 54320PRTHomo sapiensP2B-2F6 HCDR3(1)..(20)
43Ala Arg Ala Val Val Gly Ile Val Val Val Pro Ala Ala Gly Arg Arg1
5 10 15Ala Phe Asp Ile 20449PRTHomo sapiensP2B-2F6 LCDR1(1)..(9)
44Ser Ser Asp Val Gly Gly Tyr Asn Tyr1 5454PRTHomo sapiensP2B-2F6
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 45Glu Val Ser
Xaa14610PRTHomo sapiensP2B-2F6 LCDR3(1)..(10) 46Ser Ser Tyr Ala Gly
Ser Asn Asn Leu Val1 5 1047127PRTHomo sapiensP2B-2F6 VH(1)..(127)
47Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1
5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Tyr Ser Ile Ser Ser
Gly 20 25 30Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu
Glu Trp 35 40 45Ile Gly Ser Ile Tyr His Ser Gly Ser Thr Tyr Tyr Asn
Pro Ser Leu 50 55 60Lys Thr Arg Val Thr Ile Ser Val Asp Thr Ser Lys
Asn Gln Phe Ser65 70 75 80Leu Lys Leu Ser Ser Val Thr Ala Ala Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Ala Val Val Gly Ile Val Val
Val Pro Ala Ala Gly Arg Arg 100 105 110Ala Phe Asp Ile Trp Gly Gln
Gly Thr Met Val Thr Val Ser Ser 115 120 12548110PRTHomo
sapiensP2B-2F6 VL(1)..(110) 48Gln Ser Ala Leu Thr Gln Pro Pro Ser
Ala Ser Gly Ser Pro Gly Gln1 5 10 15Ser Val Thr Ile Ser Cys Thr Gly
Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn Tyr Val Ser Trp Tyr Gln
Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45Met Ile Tyr Glu Val Ser
Lys Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser Gly Ser Lys Ser
Gly Asn Thr Ala Ser Leu Thr Val Ser Gly Leu65 70 75 80Gln Ala Glu
Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Ala Gly Ser 85 90 95Asn Asn
Leu Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105
11049381DNAHomo sapiensP2B-2F6 VHnu(1)..(381) 49caggtgcagc
tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg
tctctggtta ctccatcagc agtggttact actggggctg gatccggcag
120cccccaggga aggggctgga gtggattggg agtatctatc atagtgggag
cacctactac 180aacccgtccc tcaagactcg agtcaccata tcagtagaca
cgtccaagaa ccagttctcc 240ctgaagctga gctctgtgac cgccgcagac
acggccgtct attactgtgc gagagcggtg 300gtagggattg tagtagtacc
agctgccggt cgtcgggctt ttgatatctg gggccaaggg 360acaatggtca
ccgtctcctc a 38150330DNAHomo sapiensP2B-2F6 VLnu(1)..(330)
50cagtctgccc tgactcagcc tccctccgcg tccgggtctc ctggacagtc agtcaccatc
60tcctgcactg gaaccagcag tgacgttggt ggttataact atgtctcctg gtaccaacag
120cacccaggca aagcccccaa actcatgatt tatgaggtca gtaagcggcc
ctcaggggtc 180cctgatcgct tctctggctc caagtctggc aacacggcct
ccctgaccgt ctctgggctc 240caggctgagg atgaggctga ttattactgc
agctcatatg caggcagcaa caatttggtg 300ttcggcggag ggaccaagct
gaccgtccta 330518PRTHomo sapiensP2B-2G4 HCDR1(1)..(8) 51Gly Phe Thr
Phe Ser Ser Tyr Gly1 5528PRTHomo sapiensP2B-2G4 HCDR2(1)..(8) 52Ile
Trp Tyr Asp Gly Ser Asn Lys1 55311PRTHomo sapiensP2B-2G4
HCDR3(1)..(11) 53Ala Arg Gly Ala Ala Met Val Trp Leu Asp Tyr1 5
10549PRTHomo sapiensP2B-2G4 LCDR1(1)..(9) 54Ser Ser Asp Val Gly Gly
Tyr Asn Tyr1 5554PRTHomo sapiensP2B-2G4
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 55Asp Val Ser
Xaa15611PRTHomo sapiensP2B-2G4 LCDR3(1)..(11) 56Cys Ser Tyr Ala Gly
Ser Tyr Thr Phe Val Val1 5 1057118PRTHomo sapiensP2B-2G4
VH(1)..(118) 57Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys
Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Ala Ala Met
Val Trp Leu Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val
Ser Ser 11558111PRTHomo sapiensP2B-2G4 VL(1)..(111) 58Gln Ser Ala
Leu Thr Gln Pro Arg Ser Val Ser Gly Ser Pro Gly Gln1 5 10 15Ser Val
Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn
Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40
45Met Ile Tyr Asp Val Ser Lys Arg Pro Ser Gly Val Pro Asp Arg Phe
50 55 60Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly
Leu65 70 75 80Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Cys Ser Tyr
Ala Gly Ser 85 90 95Tyr Thr Phe Val Val Phe Gly Gly Gly Thr Lys Leu
Thr Val Leu 100 105 11059354DNAHomo sapiensP2B-2G4 VHnu(1)..(354)
59caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc
60tcctgtgcag cgtctggatt caccttcagt agctatggca tgcactgggt ccgccaggct
120ccaggcaagg ggctggagtg ggtggcagtt atatggtatg atggaagtaa
taaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agccgaggac
acggctgtgt attactgtgc gagaggggca 300gctatggttt ggcttgacta
ctggggccag ggaaccctgg tcaccgtctc ctca 35460333DNAHomo
sapiensP2B-2G4 VLnu(1)..(333) 60cagtctgccc tgactcagcc tcgctcagtg
tccgggtctc ctggacagtc agtcaccatc 60tcctgcactg gaaccagcag tgatgttggt
ggttataact atgtctcctg gtaccaacag 120cacccaggca aagcccccaa
actcatgatt tatgatgtca gtaagcggcc ctcaggggtc 180cctgatcgct
tctctggctc caagtctggc aacacggcct ccctgaccat ctctgggctc
240caggctgagg atgaggctga ttattactgc tgctcatatg caggcagcta
cactttcgtg 300gtattcggcg gagggaccaa gctgaccgtc cta 33361124PRTHomo
sapiensP2B-2G11 VH(1)..(124) 61Glu Val Gln Leu Val Glu Ser Gly Gly
Gly Leu Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30Ala Met His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Gly Ile Ser Trp Asn
Gly Gly Ile Ile Gly Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr
Ile Ser Arg Asp Asn Ala Lys Thr Ser Leu Tyr65 70 75 80Leu Gln Met
Asn Ser Leu Lys Pro Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Ala Lys
Val Ala Gly Arg Gly Asp Tyr Asp Tyr Tyr Tyr Gly Met Asp 100 105
110Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115
12062111PRTHomo sapiensP2B-2G11 VL(1)..(111) 62Gln Ser Val Leu Thr
Gln Pro Pro Ser Val Ser Gly Ala Pro Gly Gln1 5 10 15Arg Val Thr Ile
Ser Cys Thr Gly Ser Ser Ser Asn Ile Gly Ala Gly 20 25 30Tyr Asp Val
His Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu 35 40 45Leu Ile
Tyr Gly Asn Asn Asn Arg Pro Ser Gly Val Pro Asp Arg Phe 50 55 60Ser
Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Thr Gly Leu65 70 75
80Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Gln Ser Tyr Asp Ser Ser
85 90 95Leu Ser Gly Ser Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu
100 105 11063372DNAHomo sapiensP2B-2G11 VHnu(1)..(372) 63gaagtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggcaggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttgat gattatgcca tgcactgggt ccggcaagct
120ccagggaagg gcctggagtg ggtctcaggt attagttgga atggtggtat
cataggctat 180gcggactctg tgaagggccg attcaccatc tccagagaca
acgccaagac ttccctgtat 240ctgcaaatga acagtctgaa acctgaggac
acggccttgt attactgtgc aaaagtcgcg 300ggaagggggg attacgacta
ctactacggt atggacgtct ggggccaagg gaccacggtc 360accgtctcct ca
37264333DNAHomo sapiensP2B-2G11 VLnu(1)..(333) 64cagtctgtgc
tgacgcagcc gccctcagtg tctggggccc cagggcagag ggtcaccatc 60tcctgcactg
ggagcagctc caacatcggg gcaggttatg atgtacactg gtaccagcaa
120cttccaggaa cagcccccaa actcctcatc tatgggaaca acaatcggcc
ctcaggggtc 180cctgaccgat tctctggctc caagtctggc acctcagcct
ccctggccat cactgggctc 240caggctgagg atgaggctga ttattactgc
cagtcctatg acagcagcct gagtggttcg 300gtattcggcg gagggaccaa
gctgaccgtc cta 333658PRTHomo sapiensP2C-1A3 HCDR1(1)..(8) 65Gly Phe
Thr Phe Ser Asp Tyr Tyr1 5668PRTHomo sapiensP2C-1A3 HCDR2(1)..(8)
66Ile Ser Ser Ser Gly Ser Thr Ile1 56712PRTHomo sapiensP2C-1A3
HCDR3(1)..(12) 67Ala Arg Asp Phe Ser His Gln Gln Leu Val Pro Ser1 5
10686PRTHomo sapiensP2C-1A3 LCDR1(1)..(6) 68Gln Gly Ile Ser Ser
Tyr1 5694PRTHomo sapiensP2C-1A3
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 69Ala Ala Ser
Xaa1709PRTHomo sapiensP2C-1A3 LCDR3(1)..(9) 70Gln Gln Leu Asn Ser
Tyr Pro Leu Thr1 571119PRTHomo sapiensP2C-1A3 VH(1)..(119) 71Gln
Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys Pro Gly Gly1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asp Tyr
20 25 30Tyr Met Ser Trp Ile Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45Ser Tyr Ile Ser Ser Ser Gly Ser Thr Ile Tyr Tyr Ala Asp
Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn
Ser Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp Phe Ser His Gln Gln Leu Val
Pro Ser Trp Gly Gln Gly 100 105 110Thr Leu Val Thr Val Ser Ser
11572107PRTHomo sapiensP2C-1A3 VL(1)..(107) 72Asp Ile Gln Leu Thr
Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr
Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr 20 25 30Leu Ala Trp
Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Ala
Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser
Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75
80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn Ser Tyr Pro Leu
85 90 95Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 100
10573357DNAHomo sapiensP2C-1A3 VHnu(1)..(357) 73caggtgcagc
tggtggagtc tgggggaggc ttggtcaagc ctggagggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt gactactaca tgagctggat ccgccaggct
120ccagggaagg ggctggagtg ggtttcatac attagtagta gtggtagtac
catatactac 180gcagactctg tgaagggccg attcaccatc tccagggaca
acgccaagaa ctcactgtat 240ctgcaaatga acagcctgag agccgaggac
acggctgtgt attactgtgc gagagatttt 300tctcatcagc agctggtacc
ttcctggggc cagggaaccc tggtcaccgt ctcctca 35774321DNAHomo
sapiensP2C-1A3 VLnu(1)..(321) 74gacatccagt tgacccagtc tccatccttc
ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc gggccagtca gggcattagc
agttatttag cctggtatca gcaaaaacca 120gggaaagccc ctaagctcct
gatctatgct gcatccactt tgcaaagtgg ggtcccatca 180aggttcagcg
gcagtggatc tgggacagaa ttcactctca caatcagcag cctgcagcct
240gaagattttg caacttatta ctgtcaacag cttaatagtt acccgctcac
tttcggcgga 300gggaccaagg tggagatcaa a 321758PRTHomo sapiensP2C-1C8
HCDR1(1)..(8) 75Gly Phe Thr Phe Arg Ser Tyr Gly1 5768PRTHomo
sapiensP2C-1C8 HCDR2(1)..(8) 76Ile Trp Tyr Asp Gly Ser Asn Lys1
57713PRTHomo sapiensP2C-1C8 HCDR3(1)..(13) 77Ala Arg Asp Ile Glu
Ile Val Val Val Asn Ile Asp Tyr1 5 107811PRTHomo sapiensP2C-1C8
LCDR1(1)..(11) 78Gln Ser Leu Val Tyr Ser Asp Gly Asn Thr Tyr1 5
10794PRTHomo sapiensP2C-1C8 LCDR2(1)..(3)misc_feature(4)..(4)Xaa is
absent. 79Lys Val Ser Xaa1809PRTHomo sapiensP2C-1C8 LCDR3(1)..(9)
80Met Gln Gly Thr His Trp Pro Tyr Thr1 581120PRTHomo sapiensP2C-1C8
VH(1)..(120) 81Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Arg Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys
Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp Ile Glu Ile
Val Val Val Asn Ile Asp Tyr Trp Gly Gln 100 105 110Gly Thr Leu Val
Thr Val Ser Ser 115 12082112PRTHomo sapiensP2C-1C8 VL(1)..(112)
82Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly1
5 10 15Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val Tyr
Ser 20 25 30Asp Gly Asn Thr Tyr Leu Asn Trp Phe Gln Gln Arg Pro Gly
Gln Ser 35 40 45Pro Arg Arg Leu Ile Tyr Lys Val Ser Ile Trp Asp Ser
Gly Val Pro 50 55 60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe
Thr Leu Lys Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val
Tyr Tyr Cys Met Gln Gly 85 90 95Thr His Trp Pro Tyr Thr Phe Gly Gln
Gly Thr Lys Leu Glu Ile Lys 100 105 11083360DNAHomo sapiensP2C-1C8
VHnu(1)..(360) 83caggtgcagc tggtggagtc tgggggaggc gtggtccagc
ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagg agctatggca
tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt
atctggtatg atggaagtaa taaatactat 180gcagactccg tgaagggccg
attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agccgaggac acggctgtgt attactgtgc gagagatata
300gagatagtag tggtaaatat tgactactgg ggccagggaa ccctggtcac
cgtctcctca 36084336DNAHomo sapiensP2C-1C8 VLnu(1)..(336)
84gatgttgtga tgactcagtc tccactctcc ctgcccgtca cccttggaca gccggcctcc
60atctcctgca ggtctagtca aagcctcgta tacagtgatg gaaacaccta cttgaattgg
120tttcagcaga ggccaggcca atctccaagg cgcctaattt ataaggtttc
tatctgggac 180tctggggtcc cagacagatt cagcggcagt gggtcaggca
ctgatttcac actgaaaatc 240agcagggtgg aggctgagga tgttggggtt
tattactgca tgcaaggtac acactggccg 300tacacttttg gccaggggac
caagctggag atcaaa 336858PRTHomo sapiensP2C-1C10 HCDR1(1)..(8) 85Gly
Gly Thr Phe Ser Ser Tyr Ala1 5868PRTHomo sapiensP2C-1C10
HCDR2(1)..(8) 86Ile Ile Pro Ile Phe Gly Thr Ala1 58711PRTHomo
sapiensP2C-1C10 HCDR3(1)..(11) 87Ala Arg Val Val Thr Gly Tyr Tyr
Phe Asp Tyr1 5 10886PRTHomo sapiensP2C-1C10 LCDR1(1)..(6) 88Gln Ser
Val Ser Ser Tyr1 5894PRTHomo sapiensP2C-1C10
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 89Asp Ala Ser
Xaa1908PRTHomo sapiensP2C-1C10 LCDR3(1)..(8) 90Gln Gln Arg Ser Asn
Trp Pro Ser1 591118PRTHomo sapiensP2C-1C10 VH(1)..(118) 91Gln Val
Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser1 5 10 15Ser
Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25
30Ala Ile Ile Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
35 40 45Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys
Phe 50 55 60Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr
Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95Ala Arg Val Val Thr Gly Tyr Tyr Phe Asp Tyr
Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val Ser Ser
11592106PRTHomo sapiensP2C-1C10 VL(1)..(106) 92Glu Ile Val Leu Thr
Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr
Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr 20 25 30Leu Ala Trp
Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 35 40 45Tyr Asp
Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 50 55 60Ser
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro65 70 75
80Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Ser
85 90 95Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100 10593354DNAHomo
sapiensP2C-1C10 VHnu(1)..(354) 93caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctgggtcctc ggtgaaggtc 60tcctgcaagg cttctggagg caccttcagc
agctatgcta tcatctgggt gcgacaggcc 120cctggacaag ggcttgagtg
gatgggaggg atcatcccta tctttggtac agcaaactac 180gcacagaagt
tccagggcag agtcacgatt accgcggacg aatccacgag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc
gagagtggta 300acggggtact actttgacta ctggggccag ggaaccctgg
tcaccgtctc ctca 35494318DNAHomo sapiensP2C-1C10 VLnu(1)..(318)
94gaaattgtgt tgacacagtc tccagccacc ctgtctttgt ctccagggga aagagccacc
60ctctcctgca gggccagtca gagtgttagc agctacttag cctggtacca acagaaacct
120ggccaggctc ccaggctcct catctatgat gcatccaaca gggccactgg
catcccagcc 180aggttcagtg gcagtgggtc tgggacagac ttcactctca
ccatcagcag cctagagcct 240gaagattttg cagtttatta ctgtcagcag
cgtagcaact ggccttcttt tggccagggg 300accaagctgg agatcaaa
318958PRTHomo sapiensP2C-1D5 HCDR1(1)..(8) 95Gly Phe Thr Phe Ser
Ser Phe Ala1 5968PRTHomo sapiensP2C-1D5 HCDR2(1)..(8) 96Ile Ser Gly
Ser Gly Gly Ser Thr1 59714PRTHomo sapiensP2C-1D5 HCDR3(1)..(14)
97Ala Lys Asp Pro Asp Gly Ser Gly Ser Trp Tyr Phe Asp Tyr1 5
10986PRTHomo sapiensP2C-1D5 LCDR1(1)..(6) 98Asn Ile Gly Ser Lys
Ser1 5994PRTHomo sapiensP2C-1D5
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 99Tyr Asp Ser
Xaa110011PRTHomo sapiensP2C-1D5 LCDR3(1)..(11) 100Gln Val Trp Asp
Ser Ser Ser Asp His His Val1 5 10101121PRTHomo sapiensP2C-1D5
VH(1)..(121) 101Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Ser Ser Phe 20 25 30Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr
Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Asp Pro Asp Gly
Ser Gly Ser Trp Tyr Phe Asp Tyr Trp Gly 100 105 110Gln Gly Thr Leu
Val Thr Val Ser Ser 115 120102108PRTHomo sapiensP2C-1D5
VL(1)..(108) 102Ser Tyr Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala
Pro Gly Lys1 5 10 15Thr Ala Arg Ile Thr Cys Gly Gly Asn Asn Ile Gly
Ser Lys Ser Val 20 25 30His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro
Val Leu Val Ile Tyr 35 40 45Tyr Asp Ser Asp Arg Pro Ser Gly Ile Pro
Glu Arg Phe Ser Gly Ser 50 55 60Asn Ser Gly Asn Thr Ala Thr Leu Thr
Ile Ser Arg Val Glu Ala Gly65 70
75 80Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser Asp
His 85 90 95His Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu 100
105103363DNAHomo sapiensP2C-1D5 VHnu(1)..(363) 103gaggtgcagc
tggtggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt cacctttagc agctttgcca tgagctgggt ccgccaggct
120ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag
cacatactac 180gcagactccg tgaagggccg gttcaccatc tccagagaca
attccaagaa cacgctgtat 240ttgcaaatga acagcctgag agccgaggac
acggccgtat attactgtgc gaaagatccg 300gatggttcgg ggagttggta
ctttgactac tggggccagg gaaccctggt caccgtctcc 360tca 363104324DNAHomo
sapiensP2C-1D5 VLnu(1)..(324) 104tcctatgtgc tgactcagcc accctcagtg
tcagtggccc caggaaagac ggccaggatt 60acctgtgggg gaaacaacat tggaagtaaa
agtgtgcact ggtaccagca gaagccaggc 120caggcccctg tgctggtcat
ctattatgat agcgaccggc cctcagggat ccctgagcga 180ttctctggct
ccaactctgg gaacaccgcc accctgacca tcagcagggt cgaagccggg
240gatgaggccg actattactg tcaggtgtgg gatagtagta gtgatcatca
tgtcttcgga 300actgggacca aggtcaccgt ccta 3241058PRTHomo
sapiensP2C-1F11 HCDR1(1)..(8) 105Gly Ile Thr Val Ser Ser Asn Tyr1
51067PRTHomo sapiensP2C-1F11 HCDR2(1)..(7) 106Ile Tyr Ser Gly Gly
Ser Thr1 510711PRTHomo sapiensP2C-1F11 HCDR3(1)..(11) 107Ala Arg
Asp Leu Val Val Tyr Gly Met Asp Val1 5 101087PRTHomo
sapiensP2C-1F11 LCDR1(1)..(7) 108Gln Ser Val Ser Ser Ser Tyr1
51094PRTHomo sapiensP2C-1F11 LCDR2(1)..(3)misc_feature(4)..(4)Xaa
is absent. 109Gly Ala Ser Xaa11108PRTHomo sapiensP2C-1F11
LCDR3(1)..(8) 110Gln Gln Tyr Gly Ser Ser Pro Thr1 5111117PRTHomo
sapiensP2C-1F11 VH(1)..(117) 111Glu Val Gln Leu Val Glu Ser Gly Gly
Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Ile Thr Val Ser Ser Asn 20 25 30Tyr Met Asn Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Leu Ile Tyr Ser Gly
Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu65 70 75 80Gln Met Asn
Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr His Cys Ala 85 90 95Arg Asp
Leu Val Val Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr 100 105
110Val Thr Val Ser Ser 115112107PRTHomo sapiensP2C-1F11
VL(1)..(107) 112Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu
Ser Pro Gly1 5 10 15Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser
Val Ser Ser Ser 20 25 30Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln
Ala Pro Arg Leu Leu 35 40 45Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly
Ile Pro Asp Arg Phe Ser 50 55 60Gly Ser Gly Ser Gly Thr Asp Phe Thr
Leu Thr Ile Ser Arg Leu Glu65 70 75 80Pro Glu Asp Phe Ala Val Tyr
Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95Thr Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys 100 105113351DNAHomo sapiensP2C-1F11
VHnu(1)..(351) 113gaggtgcagc tggtggagtc tgggggaggc ttggtccagc
ctggggggtc cctgagactc 60tcctgtgcag cctctggaat caccgtcagt agcaactaca
tgaactgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcactt
atttatagcg gtggtagcac atactacgca 180gactccgtga agggcagatt
caccatctcc agagacaatt ccaagaacac gttgtatctt 240caaatgaaca
gcctgagagc cgaggacacg gctgtgtatc actgtgcgag agatctggtg
300gtatacggta tggacgtctg gggccaaggg accacggtca ccgtctcctc a
351114321DNAHomo sapiensP2C-1F11 VLnu(1)..(321) 114gaaattgtgt
tgacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60ctctcctgca
gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa
120cctggccagg ctcccaggct cctcatctat ggtgcatcca gcagggccac
tggcatccca 180gacaggttca gtggcagtgg gtctgggaca gacttcactc
tcaccatcag cagactggag 240cctgaagatt ttgcagtgta ttactgtcag
cagtatggta gctcacccac ttttggccag 300gggaccaagc tggagatcaa a
321115330PRTHomo sapiensAmino acid sequence of heavy chain constant
region(1)..(330) 115Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala
Pro Ser Ser Lys1 5 10 15Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys
Leu Val Lys Asp Tyr 20 25 30Phe Pro Glu Pro Val Thr Val Ser Trp Asn
Ser Gly Ala Leu Thr Ser 35 40 45Gly Val His Thr Phe Pro Ala Val Leu
Gln Ser Ser Gly Leu Tyr Ser 50 55 60Leu Ser Ser Val Val Thr Val Pro
Ser Ser Ser Leu Gly Thr Gln Thr65 70 75 80Tyr Ile Cys Asn Val Asn
His Lys Pro Ser Asn Thr Lys Val Asp Lys 85 90 95Lys Val Glu Pro Lys
Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys 100 105 110Pro Ala Pro
Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro 115 120 125Lys
Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys 130 135
140Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn
Trp145 150 155 160Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr
Lys Pro Arg Glu 165 170 175Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val
Ser Val Leu Thr Val Leu 180 185 190His Gln Asp Trp Leu Asn Gly Lys
Glu Tyr Lys Cys Lys Val Ser Asn 195 200 205Lys Ala Leu Pro Ala Pro
Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly 210 215 220Gln Pro Arg Glu
Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu225 230 235 240Leu
Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr 245 250
255Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn
260 265 270Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
Phe Phe 275 280 285Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp
Gln Gln Gly Asn 290 295 300Val Phe Ser Cys Ser Val Met His Glu Ala
Leu His Asn His Tyr Thr305 310 315 320Gln Lys Ser Leu Ser Leu Ser
Pro Gly Lys 325 330116106PRTHomo sapiensAmino acid sequence of
lambda light chain constant region(1)..(106) 116Gly Gln Pro Lys Ala
Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser1 5 10 15Glu Glu Leu Gln
Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp 20 25 30Phe Tyr Pro
Gly Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro 35 40 45Val Lys
Ala Gly Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn 50 55 60Lys
Tyr Ala Ala Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys65 70 75
80Ser His Arg Ser Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val
85 90 95Glu Lys Thr Val Ala Pro Thr Glu Cys Ser 100
105117107PRTHomo sapiensAmino acid sequence of appa light chain
constant region(1)..(107) 117Arg Thr Val Ala Ala Pro Ser Val Phe
Ile Phe Pro Pro Ser Asp Glu1 5 10 15Gln Leu Lys Ser Gly Thr Ala Ser
Val Val Cys Leu Leu Asn Asn Phe 20 25 30Tyr Pro Arg Glu Ala Lys Val
Gln Trp Lys Val Asp Asn Ala Leu Gln 35 40 45Ser Gly Asn Ser Gln Glu
Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 50 55 60Thr Tyr Ser Leu Ser
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu65 70 75 80Lys His Lys
Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 85 90 95Pro Val
Thr Lys Ser Phe Asn Arg Gly Glu Cys 100 105118990DNAHomo
sapiensNucleic acid sequence of heavy chain constant
region(1)..(990) 118gcgtcgacca agggcccatc ggtcttcccc ctggcaccct
cctccaagag cacctctggg 60ggcacagcgg ccctgggctg cctggtcaag gactacttcc
ccgaacccgt gacggtgtcg 120tggaactcag gcgccctgac cagcggcgtg
cacaccttcc cggctgtcct acagtcctca 180ggactctact ccctcagcag
cgtggtgacc gtgccctcca gcagcttggg cacccagacc 240tacatctgca
acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc
300aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaact
cctgggggga 360ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc
tcatgatctc ccggacccct 420gaggtcacat gcgtggtggt ggacgtgagc
cacgaagacc ctgaggtcaa gttcaactgg 480tacgtggacg gcgtggaggt
gcataatgcc aagacaaagc cgcgggagga gcagtacaac 540agcacgtacc
gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag
600gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa
aaccatctcc 660aaagccaaag ggcagccccg agaaccacag gtgtacaccc
tgcccccatc ccgggatgag 720ctgaccaaga accaggtcag cctgacctgc
ctggtcaaag gcttctatcc cagcgacatc 780gccgtggagt gggagagcaa
tgggcagccg gagaacaact acaagaccac gcctcccgtg 840ctggactccg
acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg
900cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa
ccactacacg 960cagaagagcc tctccctgtc tccgggtaaa 990119318DNAHomo
sapiensNucleic acid sequence of lambda light chain constant
region(1)..(318) 119ggtcagccca aggctgcccc ctcggtcact ctgttcccac
cctcgagtga ggagcttcaa 60gccaacaagg ccacactggt gtgtctcata agtgacttct
acccgggagc cgtgacagtg 120gcctggaagg cagatagcag ccccgtcaag
gcgggagtgg agaccaccac accctccaaa 180caaagcaaca acaagtacgc
ggccagcagc tacctgagcc tgacgcctga gcagtggaag 240tcccacagaa
gctacagctg ccaggtcacg catgaaggga gcaccgtgga gaagacagtg
300gcccctacag aatgttca 318120321DNAHomo sapiensNucleic acid
sequence of kappa light chain constant region(1)..(321)
120cgtacggtgg ctgcaccatc tgtcttcatc ttcccgccat ctgatgagca
gttgaaatct 60ggaactgcct ctgttgtgtg cctgctgaat aacttctacc ccagagaagc
caaagtgcag 120tggaaggtgg acaacgccct gcagagcgga aacagccagg
aaagcgtgac agagcaggat 180tccaaggatt ccacatacag cctgagcagc
acactgacac tgtccaaggc cgactacgag 240aagcacaagg tgtacgcctg
cgaagtgaca caccagggac tgtcctcccc tgtgacaaag 300agcttcaaca
gaggagaatg c 3211211211PRTArtificial SequenceAmino acid sequence of
the extracellular domain of S protein of SARS-CoV-2 121Met Phe Val
Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val1 5 10 15Asn Leu
Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe 20 25 30Thr
Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu 35 40
45His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp
50 55 60Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe
Asp65 70 75 80Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala
Ser Thr Glu 85 90 95Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr
Thr Leu Asp Ser 100 105 110Lys Thr Gln Ser Leu Leu Ile Val Asn Asn
Ala Thr Asn Val Val Ile 115 120 125Lys Val Cys Glu Phe Gln Phe Cys
Asn Asp Pro Phe Leu Gly Val Tyr 130 135 140Tyr His Lys Asn Asn Lys
Ser Trp Met Glu Ser Glu Phe Arg Val Tyr145 150 155 160Ser Ser Ala
Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu 165 170 175Met
Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe 180 185
190Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr
195 200 205Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala
Leu Glu 210 215 220Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr
Arg Phe Gln Thr225 230 235 240Leu Leu Ala Leu His Arg Ser Tyr Leu
Thr Pro Gly Asp Ser Ser Ser 245 250 255Gly Trp Thr Ala Gly Ala Ala
Ala Tyr Tyr Val Gly Tyr Leu Gln Pro 260 265 270Arg Thr Phe Leu Leu
Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala 275 280 285Val Asp Cys
Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys 290 295 300Ser
Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val305 310
315 320Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu
Cys 325 330 335Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser
Val Tyr Ala 340 345 350Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala
Asp Tyr Ser Val Leu 355 360 365Tyr Asn Ser Ala Ser Phe Ser Thr Phe
Lys Cys Tyr Gly Val Ser Pro 370 375 380Thr Lys Leu Asn Asp Leu Cys
Phe Thr Asn Val Tyr Ala Asp Ser Phe385 390 395 400Val Ile Arg Gly
Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly 405 410 415Lys Ile
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys 420 425
430Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn
435 440 445Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys
Pro Phe 450 455 460Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly
Ser Thr Pro Cys465 470 475 480Asn Gly Val Glu Gly Phe Asn Cys Tyr
Phe Pro Leu Gln Ser Tyr Gly 485 490 495Phe Gln Pro Thr Asn Gly Val
Gly Tyr Gln Pro Tyr Arg Val Val Val 500 505 510Leu Ser Phe Glu Leu
Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys 515 520 525Lys Ser Thr
Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn 530 535 540Gly
Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu545 550
555 560Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala
Val 565 570 575Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro
Cys Ser Phe 580 585 590Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn
Thr Ser Asn Gln Val 595 600 605Ala Val Leu Tyr Gln Asp Val Asn Cys
Thr Glu Val Pro Val Ala Ile 610 615 620His Ala Asp Gln Leu Thr Pro
Thr Trp Arg Val Tyr Ser Thr Gly Ser625 630 635 640Asn Val Phe Gln
Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val 645 650 655Asn Asn
Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala 660 665
670Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala
675 680 685Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu
Asn Ser 690 695 700Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr
Asn Phe Thr Ile705 710 715 720Ser Val Thr Thr Glu Ile Leu Pro Val
Ser Met Thr Lys Thr Ser Val 725 730 735Asp Cys Thr Met Tyr Ile Cys
Gly Asp Ser Thr Glu Cys Ser Asn Leu 740 745 750Leu Leu Gln Tyr Gly
Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 755 760 765Gly Ile Ala
Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 770 775 780Val
Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe785 790
795 800Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg
Ser 805 810 815Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala
Asp Ala Gly 820 825 830Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp
Ile Ala Ala Arg Asp 835 840 845Leu Ile Cys Ala Gln Lys Phe Asn Gly
Leu Thr Val Leu Pro Pro Leu 850 855 860Leu Thr Asp Glu Met Ile Ala
Gln Tyr Thr Ser Ala Leu Leu Ala Gly865 870 875 880Thr Ile Thr Ser
Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 885 890 895Pro Phe
Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 900 905
910Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn
915 920
925Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala
930 935 940Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala
Leu Asn945 950 955 960Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly
Ala Ile Ser Ser Val 965 970 975Leu Asn Asp Ile Leu Ser Arg Leu Asp
Lys Val Glu Ala Glu Val Gln 980 985 990Ile Asp Arg Leu Ile Thr Gly
Arg Leu Gln Ser Leu Gln Thr Tyr Val 995 1000 1005Thr Gln Gln Leu
Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn 1010 1015 1020Leu Ala
Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys 1025 1030
1035Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro
1040 1045 1050Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr
Tyr Val 1055 1060 1065Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro
Ala Ile Cys His 1070 1075 1080Asp Gly Lys Ala His Phe Pro Arg Glu
Gly Val Phe Val Ser Asn 1085 1090 1095Gly Thr His Trp Phe Val Thr
Gln Arg Asn Phe Tyr Glu Pro Gln 1100 1105 1110Ile Ile Thr Thr Asp
Asn Thr Phe Val Ser Gly Asn Cys Asp Val 1115 1120 1125Val Ile Gly
Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro 1130 1135 1140Glu
Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn 1145 1150
1155His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn
1160 1165 1170Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu
Asn Glu 1175 1180 1185Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp
Leu Gln Glu Leu 1190 1195 1200Gly Lys Tyr Glu Gln Tyr Ile Lys 1205
12101221195PRTArtificial SequenceAmino acid sequence of the
extracellular domain of S protein of SARS-CoV 122Met Phe Ile Phe
Leu Leu Phe Leu Thr Leu Thr Ser Gly Ser Asp Leu1 5 10 15Asp Arg Cys
Thr Thr Phe Asp Asp Val Gln Ala Pro Asn Tyr Thr Gln 20 25 30His Thr
Ser Ser Met Arg Gly Val Tyr Tyr Pro Asp Glu Ile Phe Arg 35 40 45Ser
Asp Thr Leu Tyr Leu Thr Gln Asp Leu Phe Leu Pro Phe Tyr Ser 50 55
60Asn Val Thr Gly Phe His Thr Ile Asn His Thr Phe Gly Asn Pro Val65
70 75 80Ile Pro Phe Lys Asp Gly Ile Tyr Phe Ala Ala Thr Glu Lys Ser
Asn 85 90 95Val Val Arg Gly Trp Val Phe Gly Ser Thr Met Asn Asn Lys
Ser Gln 100 105 110Ser Val Ile Ile Ile Asn Asn Ser Thr Asn Val Val
Ile Arg Ala Cys 115 120 125Asn Phe Glu Leu Cys Asp Asn Pro Phe Phe
Ala Val Ser Lys Pro Met 130 135 140Gly Thr Gln Thr His Thr Met Ile
Phe Asp Asn Ala Phe Asn Cys Thr145 150 155 160Phe Glu Tyr Ile Ser
Asp Ala Phe Ser Leu Asp Val Ser Glu Lys Ser 165 170 175Gly Asn Phe
Lys His Leu Arg Glu Phe Val Phe Lys Asn Lys Asp Gly 180 185 190Phe
Leu Tyr Val Tyr Lys Gly Tyr Gln Pro Ile Asp Val Val Arg Asp 195 200
205Leu Pro Ser Gly Phe Asn Thr Leu Lys Pro Ile Phe Lys Leu Pro Leu
210 215 220Gly Ile Asn Ile Thr Asn Phe Arg Ala Ile Leu Thr Ala Phe
Ser Pro225 230 235 240Ala Gln Asp Ile Trp Gly Thr Ser Ala Ala Ala
Tyr Phe Val Gly Tyr 245 250 255Leu Lys Pro Thr Thr Phe Met Leu Lys
Tyr Asp Glu Asn Gly Thr Ile 260 265 270Thr Asp Ala Val Asp Cys Ser
Gln Asn Pro Leu Ala Glu Leu Lys Cys 275 280 285Ser Val Lys Ser Phe
Glu Ile Asp Lys Gly Ile Tyr Gln Thr Ser Asn 290 295 300Phe Arg Val
Val Pro Ser Gly Asp Val Val Arg Phe Pro Asn Ile Thr305 310 315
320Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Lys Phe Pro Ser
325 330 335Val Tyr Ala Trp Glu Arg Lys Lys Ile Ser Asn Cys Val Ala
Asp Tyr 340 345 350Ser Val Leu Tyr Asn Ser Thr Phe Phe Ser Thr Phe
Lys Cys Tyr Gly 355 360 365Val Ser Ala Thr Lys Leu Asn Asp Leu Cys
Phe Ser Asn Val Tyr Ala 370 375 380Asp Ser Phe Val Val Lys Gly Asp
Asp Val Arg Gln Ile Ala Pro Gly385 390 395 400Gln Thr Gly Val Ile
Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe 405 410 415Met Gly Cys
Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr Ser 420 425 430Thr
Gly Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg His Gly Lys Leu 435 440
445Arg Pro Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp Gly
450 455 460Lys Pro Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp Pro Leu
Asn Asp465 470 475 480Tyr Gly Phe Tyr Thr Thr Thr Gly Ile Gly Tyr
Gln Pro Tyr Arg Val 485 490 495Val Val Leu Ser Phe Glu Leu Leu Asn
Ala Pro Ala Thr Val Cys Gly 500 505 510Pro Lys Leu Ser Thr Asp Leu
Ile Lys Asn Gln Cys Val Asn Phe Asn 515 520 525Phe Asn Gly Leu Thr
Gly Thr Gly Val Leu Thr Pro Ser Ser Lys Arg 530 535 540Phe Gln Pro
Phe Gln Gln Phe Gly Arg Asp Val Ser Asp Phe Thr Asp545 550 555
560Ser Val Arg Asp Pro Lys Thr Ser Glu Ile Leu Asp Ile Ser Pro Cys
565 570 575Ala Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Ala
Ser Ser 580 585 590Glu Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr
Asp Val Ser Thr 595 600 605Ala Ile His Ala Asp Gln Leu Thr Pro Ala
Trp Arg Ile Tyr Ser Thr 610 615 620Gly Asn Asn Val Phe Gln Thr Gln
Ala Gly Cys Leu Ile Gly Ala Glu625 630 635 640His Val Asp Thr Ser
Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile 645 650 655Cys Ala Ser
Tyr His Thr Val Ser Leu Leu Arg Ser Thr Ser Gln Lys 660 665 670Ser
Ile Val Ala Tyr Thr Met Ser Leu Gly Ala Asp Ser Ser Ile Ala 675 680
685Tyr Ser Asn Asn Thr Ile Ala Ile Pro Thr Asn Phe Ser Ile Ser Ile
690 695 700Thr Thr Glu Val Met Pro Val Ser Met Ala Lys Thr Ser Val
Asp Cys705 710 715 720Asn Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys
Ala Asn Leu Leu Leu 725 730 735Gln Tyr Gly Ser Phe Cys Thr Gln Leu
Asn Arg Ala Leu Ser Gly Ile 740 745 750Ala Ala Glu Gln Asp Arg Asn
Thr Arg Glu Val Phe Ala Gln Val Lys 755 760 765Gln Met Tyr Lys Thr
Pro Thr Leu Lys Tyr Phe Gly Gly Phe Asn Phe 770 775 780Ser Gln Ile
Leu Pro Asp Pro Leu Lys Pro Thr Lys Arg Ser Phe Ile785 790 795
800Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly Phe Met
805 810 815Lys Gln Tyr Gly Glu Cys Leu Gly Asp Ile Asn Ala Arg Asp
Leu Ile 820 825 830Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro
Pro Leu Leu Thr 835 840 845Asp Asp Met Ile Ala Ala Tyr Thr Ala Ala
Leu Val Ser Gly Thr Ala 850 855 860Thr Ala Gly Trp Thr Phe Gly Ala
Gly Ala Ala Leu Gln Ile Pro Phe865 870 875 880Ala Met Gln Met Ala
Tyr Arg Phe Asn Gly Ile Gly Val Thr Gln Asn 885 890 895Val Leu Tyr
Glu Asn Gln Lys Gln Ile Ala Asn Gln Phe Asn Lys Ala 900 905 910Ile
Ser Gln Ile Gln Glu Ser Leu Thr Thr Thr Ser Thr Ala Leu Gly 915 920
925Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn Thr Leu
930 935 940Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val
Leu Asn945 950 955 960Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala
Glu Val Gln Ile Asp 965 970 975Arg Leu Ile Thr Gly Arg Leu Gln Ser
Leu Gln Thr Tyr Val Thr Gln 980 985 990Gln Leu Ile Arg Ala Ala Glu
Ile Arg Ala Ser Ala Asn Leu Ala Ala 995 1000 1005Thr Lys Met Ser
Glu Cys Val Leu Gly Gln Ser Lys Arg Val Asp 1010 1015 1020Phe Cys
Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln Ala Ala 1025 1030
1035Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ser Gln
1040 1045 1050Glu Arg Asn Phe Thr Thr Ala Pro Ala Ile Cys His Glu
Gly Lys 1055 1060 1065Ala Tyr Phe Pro Arg Glu Gly Val Phe Val Phe
Asn Gly Thr Ser 1070 1075 1080Trp Phe Ile Thr Gln Arg Asn Phe Phe
Ser Pro Gln Ile Ile Thr 1085 1090 1095Thr Asp Asn Thr Phe Val Ser
Gly Asn Cys Asp Val Val Ile Gly 1100 1105 1110Ile Ile Asn Asn Thr
Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp 1115 1120 1125Ser Phe Lys
Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser 1130 1135 1140Pro
Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val 1145 1150
1155Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys
1160 1165 1170Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly
Lys Tyr 1175 1180 1185Glu Gln Tyr Ile Lys Trp Pro 1190
11951231290PRTArtificial SequenceAmino acid sequence of the
extracellular domain of S protein of MERS-CoV 123Met Ile His Ser
Val Phe Leu Leu Met Phe Leu Leu Thr Pro Thr Glu1 5 10 15Ser Tyr Val
Asp Val Gly Pro Asp Ser Val Lys Ser Ala Cys Ile Glu 20 25 30Val Asp
Ile Gln Gln Thr Phe Phe Asp Lys Thr Trp Pro Arg Pro Ile 35 40 45Asp
Val Ser Lys Ala Asp Gly Ile Ile Tyr Pro Gln Gly Arg Thr Tyr 50 55
60Ser Asn Ile Thr Ile Thr Tyr Gln Gly Leu Phe Pro Tyr Gln Gly Asp65
70 75 80His Gly Asp Met Tyr Val Tyr Ser Ala Gly His Ala Thr Gly Thr
Thr 85 90 95Pro Gln Lys Leu Phe Val Ala Asn Tyr Ser Gln Asp Val Lys
Gln Phe 100 105 110Ala Asn Gly Phe Val Val Arg Ile Gly Ala Ala Ala
Asn Ser Thr Gly 115 120 125Thr Val Ile Ile Ser Pro Ser Thr Ser Ala
Thr Ile Arg Lys Ile Tyr 130 135 140Pro Ala Phe Met Leu Gly Ser Ser
Val Gly Asn Phe Ser Asp Gly Lys145 150 155 160Met Gly Arg Phe Phe
Asn His Thr Leu Val Leu Leu Pro Asp Gly Cys 165 170 175Gly Thr Leu
Leu Arg Ala Phe Tyr Cys Ile Leu Glu Pro Arg Ser Gly 180 185 190Asn
His Cys Pro Ala Gly Asn Ser Tyr Thr Ser Phe Ala Thr Tyr His 195 200
205Thr Pro Ala Thr Asp Cys Ser Asp Gly Asn Tyr Asn Arg Asn Ala Ser
210 215 220Leu Asn Ser Phe Lys Glu Tyr Phe Asn Leu Arg Asn Cys Thr
Phe Met225 230 235 240Tyr Thr Tyr Asn Ile Thr Glu Asp Glu Ile Leu
Glu Trp Phe Gly Ile 245 250 255Thr Gln Thr Ala Gln Gly Val His Leu
Phe Ser Ser Arg Tyr Val Asp 260 265 270Leu Tyr Gly Gly Asn Met Phe
Gln Phe Ala Thr Leu Pro Val Tyr Asp 275 280 285Thr Ile Lys Tyr Tyr
Ser Ile Ile Pro His Ser Ile Arg Ser Ile Gln 290 295 300Ser Asp Arg
Lys Ala Trp Ala Ala Phe Tyr Val Tyr Lys Leu Gln Pro305 310 315
320Leu Thr Phe Leu Leu Asp Phe Ser Val Asp Gly Tyr Ile Arg Arg Ala
325 330 335Ile Asp Cys Gly Phe Asn Asp Leu Ser Gln Leu His Cys Ser
Tyr Glu 340 345 350Ser Phe Asp Val Glu Ser Gly Val Tyr Ser Val Ser
Ser Phe Glu Ala 355 360 365Lys Pro Ser Gly Ser Val Val Glu Gln Ala
Glu Gly Val Glu Cys Asp 370 375 380Phe Ser Pro Leu Leu Ser Gly Thr
Pro Pro Gln Val Tyr Asn Phe Lys385 390 395 400Arg Leu Val Phe Thr
Asn Cys Asn Tyr Asn Leu Thr Lys Leu Leu Ser 405 410 415Leu Phe Ser
Val Asn Asp Phe Thr Cys Ser Gln Ile Ser Pro Ala Ala 420 425 430Ile
Ala Ser Asn Cys Tyr Ser Ser Leu Ile Leu Asp Tyr Phe Ser Tyr 435 440
445Pro Leu Ser Met Lys Ser Asp Leu Ser Val Ser Ser Ala Gly Pro Ile
450 455 460Ser Gln Phe Asn Tyr Lys Gln Ser Phe Ser Asn Pro Thr Cys
Leu Ile465 470 475 480Leu Ala Thr Val Pro His Asn Leu Thr Thr Ile
Thr Lys Pro Leu Lys 485 490 495Tyr Ser Tyr Ile Asn Lys Cys Ser Arg
Leu Leu Ser Asp Asp Arg Thr 500 505 510Glu Val Pro Gln Leu Val Asn
Ala Asn Gln Tyr Ser Pro Cys Val Ser 515 520 525Ile Val Pro Ser Thr
Val Trp Glu Asp Gly Asp Tyr Tyr Arg Lys Gln 530 535 540Leu Ser Pro
Leu Glu Gly Gly Gly Trp Leu Val Ala Ser Gly Ser Thr545 550 555
560Val Ala Met Thr Glu Gln Leu Gln Met Gly Phe Gly Ile Thr Val Gln
565 570 575Tyr Gly Thr Asp Thr Asn Ser Val Cys Pro Lys Leu Glu Phe
Ala Asn 580 585 590Asp Thr Lys Ile Ala Ser Gln Leu Gly Asn Cys Val
Glu Tyr Ser Leu 595 600 605Tyr Gly Val Ser Gly Arg Gly Val Phe Gln
Asn Cys Thr Ala Val Gly 610 615 620Val Arg Gln Gln Arg Phe Val Tyr
Asp Ala Tyr Gln Asn Leu Val Gly625 630 635 640Tyr Tyr Ser Asp Asp
Gly Asn Tyr Tyr Cys Leu Arg Ala Cys Val Ser 645 650 655Val Pro Val
Ser Val Ile Tyr Asp Lys Glu Thr Lys Thr His Ala Thr 660 665 670Leu
Phe Gly Ser Val Ala Cys Glu His Ile Ser Ser Thr Met Ser Gln 675 680
685Tyr Ser Arg Ser Thr Arg Ser Met Leu Lys Arg Arg Asp Ser Thr Tyr
690 695 700Gly Pro Leu Gln Thr Pro Val Gly Cys Val Leu Gly Leu Val
Asn Ser705 710 715 720Ser Leu Phe Val Glu Asp Cys Lys Leu Pro Leu
Gly Gln Ser Leu Cys 725 730 735Ala Leu Pro Asp Thr Pro Ser Thr Leu
Thr Pro Arg Ser Val Arg Ser 740 745 750Val Pro Gly Glu Met Arg Leu
Ala Ser Ile Ala Phe Asn His Pro Ile 755 760 765Gln Val Asp Gln Leu
Asn Ser Ser Tyr Phe Lys Leu Ser Ile Pro Thr 770 775 780Asn Phe Ser
Phe Gly Val Thr Gln Glu Tyr Ile Gln Thr Thr Ile Gln785 790 795
800Lys Val Thr Val Asp Cys Lys Gln Tyr Val Cys Asn Gly Phe Gln Lys
805 810 815Cys Glu Gln Leu Leu Arg Glu Tyr Gly Gln Phe Cys Ser Lys
Ile Asn 820 825 830Gln Ala Leu His Gly Ala Asn Leu Arg Gln Asp Asp
Ser Val Arg Asn 835 840 845Leu Phe Ala Ser Val Lys Ser Ser Gln Ser
Ser Pro Ile Ile Pro Gly 850 855 860Phe Gly Gly Asp Phe Asn Leu Thr
Leu Leu Glu Pro Val Ser Ile Ser865 870 875 880Thr Gly Ser Arg Ser
Ala Arg Ser Ala Ile Glu Asp Leu Leu Phe Asp 885 890 895Lys Val Thr
Ile Ala Asp Pro Gly Tyr Met Gln Gly Tyr Asp Asp Cys 900 905 910Met
Gln Gln Gly Pro Ala Ser Ala Arg Asp Leu Ile Cys Ala Gln Tyr 915 920
925Val Ala Gly Tyr Lys Val Leu Pro Pro Leu Met Asp Val Asn Met Glu
930 935 940Ala Ala Tyr Thr Ser Ser Leu Leu Gly Ser Ile Ala Gly Val
Gly Trp945 950
955 960Thr Ala Gly Leu Ser Ser Phe Ala Ala Ile Pro Phe Ala Gln Ser
Ile 965 970 975Phe Tyr Arg Leu Asn Gly Val Gly Ile Thr Gln Gln Val
Leu Ser Glu 980 985 990Asn Gln Lys Leu Ile Ala Asn Lys Phe Asn Gln
Ala Leu Gly Ala Met 995 1000 1005Gln Thr Gly Phe Thr Thr Thr Asn
Glu Ala Phe Gln Lys Val Gln 1010 1015 1020Asp Ala Val Asn Asn Asn
Ala Gln Ala Leu Ser Lys Leu Ala Ser 1025 1030 1035Glu Leu Ser Asn
Thr Phe Gly Ala Ile Ser Ala Ser Ile Gly Asp 1040 1045 1050Ile Ile
Gln Arg Leu Asp Val Leu Glu Gln Asp Ala Gln Ile Asp 1055 1060
1065Arg Leu Ile Asn Gly Arg Leu Thr Thr Leu Asn Ala Phe Val Ala
1070 1075 1080Gln Gln Leu Val Arg Ser Glu Ser Ala Ala Leu Ser Ala
Gln Leu 1085 1090 1095Ala Lys Asp Lys Val Asn Glu Cys Val Lys Ala
Gln Ser Lys Arg 1100 1105 1110Ser Gly Phe Cys Gly Gln Gly Thr His
Ile Val Ser Phe Val Val 1115 1120 1125Asn Ala Pro Asn Gly Leu Tyr
Phe Met His Val Gly Tyr Tyr Pro 1130 1135 1140Ser Asn His Ile Glu
Val Val Ser Ala Tyr Gly Leu Cys Asp Ala 1145 1150 1155Ala Asn Pro
Thr Asn Cys Ile Ala Pro Val Asn Gly Tyr Phe Ile 1160 1165 1170Lys
Thr Asn Asn Thr Arg Ile Val Asp Glu Trp Ser Tyr Thr Gly 1175 1180
1185Ser Ser Phe Tyr Ala Pro Glu Pro Ile Thr Ser Leu Asn Thr Lys
1190 1195 1200Tyr Val Ala Pro Gln Val Thr Tyr Gln Asn Ile Ser Thr
Asn Leu 1205 1210 1215Pro Pro Pro Leu Leu Gly Asn Ser Thr Gly Ile
Asp Phe Gln Asp 1220 1225 1230Glu Leu Asp Glu Phe Phe Lys Asn Val
Ser Thr Ser Ile Pro Asn 1235 1240 1245Phe Gly Ser Leu Thr Gln Ile
Asn Thr Thr Leu Leu Asp Leu Thr 1250 1255 1260Tyr Glu Met Leu Ser
Leu Gln Gln Val Val Lys Ala Leu Asn Glu 1265 1270 1275Ser Tyr Ile
Asp Leu Lys Glu Leu Gly Asn Tyr Thr 1280 1285
1290124222PRTArtificial SequenceAmino acid sequence of the spike
protein RBD of SARS-CoV 124Arg Val Val Pro Ser Gly Asp Val Val Arg
Phe Pro Asn Ile Thr Asn1 5 10 15Leu Cys Pro Phe Gly Glu Val Phe Asn
Ala Thr Lys Phe Pro Ser Val 20 25 30Tyr Ala Trp Glu Arg Lys Lys Ile
Ser Asn Cys Val Ala Asp Tyr Ser 35 40 45Val Leu Tyr Asn Ser Thr Phe
Phe Ser Thr Phe Lys Cys Tyr Gly Val 50 55 60Ser Ala Thr Lys Leu Asn
Asp Leu Cys Phe Ser Asn Val Tyr Ala Asp65 70 75 80Ser Phe Val Val
Lys Gly Asp Asp Val Arg Gln Ile Ala Pro Gly Gln 85 90 95Thr Gly Val
Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Met 100 105 110Gly
Cys Val Leu Ala Trp Asn Thr Arg Asn Ile Asp Ala Thr Ser Thr 115 120
125Gly Asn Tyr Asn Tyr Lys Tyr Arg Tyr Leu Arg His Gly Lys Leu Arg
130 135 140Pro Phe Glu Arg Asp Ile Ser Asn Val Pro Phe Ser Pro Asp
Gly Lys145 150 155 160Pro Cys Thr Pro Pro Ala Leu Asn Cys Tyr Trp
Pro Leu Asn Asp Tyr 165 170 175Gly Phe Tyr Thr Thr Thr Gly Ile Gly
Tyr Gln Pro Tyr Arg Val Val 180 185 190Val Leu Ser Phe Glu Leu Leu
Asn Ala Pro Ala Thr Val Cys Gly Pro 195 200 205Lys Leu Ser Thr Asp
Leu Ile Lys Asn Gln Cys Val Asn Phe 210 215 220125666DNAArtificial
SequenceNucleic acid sequence of the spike protein RBD of SARS-CoV
125cgggtggtgc ccagcggcga cgtggtgcgg ttccccaaca tcaccaacct
gtgccccttc 60ggcgaggtgt tcaacgccac caagttcccc agcgtgtacg cctgggagcg
gaagaagatc 120agcaactgcg tggccgacta cagcgtgctg tacaacagca
ccttcttcag caccttcaag 180tgctacggcg tgagcgccac caagctgaac
gacctgtgct tcagcaacgt gtacgccgac 240agcttcgtgg tgaagggcga
cgacgtgcgg cagatcgccc ccggccagac cggcgtgatc 300gccgactaca
actacaagct gcccgacgac ttcatgggct gcgtgctggc ctggaacacc
360cggaacatcg acgccaccag caccggcaac tacaactaca agtaccggta
cctgcggcac 420ggcaagctgc ggcccttcga gcgggacatc agcaacgtgc
ccttcagccc cgacggcaag 480ccctgcaccc cccccgccct gaactgctac
tggcccctga acgactacgg cttctacacc 540actaccggca tcggctacca
gccctaccgg gtggtggtgc tgagcttcga gctgctgaac 600gcccccgcca
ccgtgtgcgg ccccaagctg agcaccgacc tgatcaagaa ccagtgcgtg 660aacttc
666126208PRTArtificial SequenceAmino acid sequence of the spike
protein RBD of MERS-CoV 126Val Glu Cys Asp Phe Ser Pro Leu Leu Ser
Gly Thr Pro Pro Gln Val1 5 10 15Tyr Asn Phe Lys Arg Leu Val Phe Thr
Asn Cys Asn Tyr Asn Leu Thr 20 25 30Lys Leu Leu Ser Leu Phe Ser Val
Asn Asp Phe Thr Cys Ser Gln Ile 35 40 45Ser Pro Ala Ala Ile Ala Ser
Asn Cys Tyr Ser Ser Leu Ile Leu Asp 50 55 60Tyr Phe Ser Tyr Pro Leu
Ser Met Lys Ser Asp Leu Ser Val Ser Ser65 70 75 80Ala Gly Pro Ile
Ser Gln Phe Asn Tyr Lys Gln Ser Phe Ser Asn Pro 85 90 95Thr Cys Leu
Ile Leu Ala Thr Val Pro His Asn Leu Thr Thr Ile Thr 100 105 110Lys
Pro Leu Lys Tyr Ser Tyr Ile Asn Lys Cys Ser Arg Leu Leu Ser 115 120
125Asp Asp Arg Thr Glu Val Pro Gln Leu Val Asn Ala Asn Gln Tyr Ser
130 135 140Pro Cys Val Ser Ile Val Pro Ser Thr Val Trp Glu Asp Gly
Asp Tyr145 150 155 160Tyr Arg Lys Gln Leu Ser Pro Leu Glu Gly Gly
Gly Trp Leu Val Ala 165 170 175Ser Gly Ser Thr Val Ala Met Thr Glu
Gln Leu Gln Met Gly Phe Gly 180 185 190Ile Thr Val Gln Tyr Gly Thr
Asp Thr Asn Ser Val Cys Pro Lys Leu 195 200 205127624DNAArtificial
SequenceNucleic acid sequence of the spike protein RBD of MERS-CoV
127gtggagtgtg acttcagccc actgctgtct ggcacacctc cacaggtcta
caacttcaag 60agactggtgt tcaccaactg taactacaac ctgaccaaac tgctgtccct
gttctctgtg 120aatgacttca cttgtagcca gattagccct gctgccattg
ccagcaactg ttactcctcc 180ctgattctgg actacttctc ctacccactg
agtatgaagt ctgacctgtc tgtgtcctct 240gctggaccaa tcagccagtt
caactacaag cagtccttca gcaacccaac ttgtctgatt 300ctggctacag
tgccacacaa cctgaccacc atcaccaagc cactgaaata ctcctacatc
360aacaagtgta gcagactgct gtctgatgac aggacagagg tgccacaact
agtgaatgcc 420aaccaataca gcccatgtgt gagcattgtg ccaagcacag
tgtgggagga tggagactac 480tacaggaagc aacttagccc attggaggga
ggaggctggc tggtggcatc tggcagcaca 540gtggctatga cagaacaact
ccaaatgggc tttggcatca cagtccaata tggcacagac 600accaactctg
tgtgtccaaa attg 624128223PRTArtificial SequenceAmino acid sequence
of the spike protein RBD of SARS-CoV-2 128Arg Val Gln Pro Thr Glu
Ser Ile Val Arg Phe Pro Asn Ile Thr Asn1 5 10 15Leu Cys Pro Phe Gly
Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val 20 25 30Tyr Ala Trp Asn
Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser 35 40 45Val Leu Tyr
Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val 50 55 60Ser Pro
Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp65 70 75
80Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln
85 90 95Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe
Thr 100 105 110Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser
Lys Val Gly 115 120 125Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg
Lys Ser Asn Leu Lys 130 135 140Pro Phe Glu Arg Asp Ile Ser Thr Glu
Ile Tyr Gln Ala Gly Ser Thr145 150 155 160Pro Cys Asn Gly Val Glu
Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser 165 170 175Tyr Gly Phe Gln
Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val 180 185 190Val Val
Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly 195 200
205Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe 210
215 220129669DNAArtificial SequenceNucleic acid sequence of the
spike protein RBD of SARS-CoV-2 129cgcgtgcagc ccaccgagag catcgtgcgc
ttccccaaca tcaccaacct gtgccccttc 60ggcgaggtgt tcaacgccac ccgcttcgcc
agcgtgtacg cctggaaccg caagcgcatc 120agcaactgcg tggccgacta
cagcgtgctg tacaacagcg ccagcttcag caccttcaag 180tgctacggcg
tgagccccac caagctgaac gacctgtgct tcaccaacgt gtacgccgac
240agcttcgtga tccgcggcga cgaggtgcgc cagatcgccc ccggccagac
cggcaagatc 300gccgactaca actacaagct gcccgacgac ttcaccggct
gcgtgatcgc ctggaacagc 360aacaacctgg acagcaaggt gggcggcaac
tacaactacc tgtaccgcct gttccgcaag 420agcaacctga agcccttcga
gcgcgacatc agcaccgaga tctaccaggc cggcagcacc 480ccctgcaacg
gcgtggaggg cttcaactgc tacttccccc tgcagagcta cggcttccag
540cccaccaacg gcgtgggcta ccagccctac cgcgtggtgg tgctgagctt
cgagctgctg 600cacgcccccg ccaccgtgtg cggccccaag aagagcacca
acctggtgaa gaacaagtgc 660gtgaacttc 66913019PRTArtificial
SequenceSignal peptide 130Met Gly Trp Ser Cys Ile Ile Leu Phe Leu
Val Ala Thr Ala Thr Gly1 5 10 15Val His Ser13119PRTArtificial
SequenceSignal peptide 131Met Gly Trp Ser Cys Ile Ile Leu Phe Leu
Val Ala Thr Ala Thr Gly1 5 10 15Ser Trp Ala1326PRTArtificial
Sequence6XHis Tag 132His His His His His His1 513310PRTArtificial
SequenceTag 133Met Ser Tyr Tyr His His His His His His1 5
101341281PRTArtificial SequenceAmino acid sequence of full length S
protein of SARS-CoV-2 134Met Phe Val Phe Leu Val Leu Leu Pro Leu
Val Ser Ser Gln Cys Val1 5 10 15Asn Leu Thr Thr Arg Thr Gln Leu Pro
Pro Ala Tyr Thr Asn Ser Phe 20 25 30Thr Arg Gly Val Tyr Tyr Pro Asp
Lys Val Phe Arg Ser Ser Val Leu 35 40 45His Ser Thr Gln Asp Leu Phe
Leu Pro Phe Phe Ser Asn Val Thr Trp 50 55 60Phe His Ala Ile His Val
Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp65 70 75 80Asn Pro Val Leu
Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu 85 90 95Lys Ser Asn
Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser 100 105 110Lys
Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile 115 120
125Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr
130 135 140Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg
Val Tyr145 150 155 160Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val
Ser Gln Pro Phe Leu 165 170 175Met Asp Leu Glu Gly Lys Gln Gly Asn
Phe Lys Asn Leu Arg Glu Phe 180 185 190Val Phe Lys Asn Ile Asp Gly
Tyr Phe Lys Ile Tyr Ser Lys His Thr 195 200 205Pro Ile Asn Leu Val
Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu 210 215 220Pro Leu Val
Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr225 230 235
240Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser
245 250 255Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu
Gln Pro 260 265 270Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr
Ile Thr Asp Ala 275 280 285Val Asp Cys Ala Leu Asp Pro Leu Ser Glu
Thr Lys Cys Thr Leu Lys 290 295 300Ser Phe Thr Val Glu Lys Gly Ile
Tyr Gln Thr Ser Asn Phe Arg Val305 310 315 320Gln Pro Thr Glu Ser
Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys 325 330 335Pro Phe Gly
Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala 340 345 350Trp
Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu 355 360
365Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro
370 375 380Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp
Ser Phe385 390 395 400Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala
Pro Gly Gln Thr Gly 405 410 415Lys Ile Ala Asp Tyr Asn Tyr Lys Leu
Pro Asp Asp Phe Thr Gly Cys 420 425 430Val Ile Ala Trp Asn Ser Asn
Asn Leu Asp Ser Lys Val Gly Gly Asn 435 440 445Tyr Asn Tyr Leu Tyr
Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe 450 455 460Glu Arg Asp
Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys465 470 475
480Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly
485 490 495Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val
Val Val 500 505 510Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val
Cys Gly Pro Lys 515 520 525Lys Ser Thr Asn Leu Val Lys Asn Lys Cys
Val Asn Phe Asn Phe Asn 530 535 540Gly Leu Thr Gly Thr Gly Val Leu
Thr Glu Ser Asn Lys Lys Phe Leu545 550 555 560Pro Phe Gln Gln Phe
Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val 565 570 575Arg Asp Pro
Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe 580 585 590Gly
Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val 595 600
605Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile
610 615 620His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr
Gly Ser625 630 635 640Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile
Gly Ala Glu His Val 645 650 655Asn Asn Ser Tyr Glu Cys Asp Ile Pro
Ile Gly Ala Gly Ile Cys Ala 660 665 670Ser Tyr Gln Thr Gln Thr Asn
Ser Pro Arg Arg Ala Arg Ser Val Ala 675 680 685Ser Gln Ser Ile Ile
Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 690 695 700Val Ala Tyr
Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile705 710 715
720Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val
725 730 735Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser
Asn Leu 740 745 750Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn
Arg Ala Leu Thr 755 760 765Gly Ile Ala Val Glu Gln Asp Lys Asn Thr
Gln Glu Val Phe Ala Gln 770 775 780Val Lys Gln Ile Tyr Lys Thr Pro
Pro Ile Lys Asp Phe Gly Gly Phe785 790 795 800Asn Phe Ser Gln Ile
Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 805 810 815Phe Ile Glu
Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 820 825 830Phe
Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 835 840
845Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu
850 855 860Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu
Ala Gly865 870 875 880Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly
Ala Ala Leu Gln Ile 885 890 895Pro Phe Ala Met Gln Met Ala Tyr Arg
Phe Asn Gly Ile Gly Val Thr 900 905 910Gln Asn Val Leu Tyr Glu Asn
Gln Lys Leu Ile Ala Asn Gln Phe Asn 915 920 925Ser Ala Ile Gly Lys
Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 930 935 940Leu Gly Lys
Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn945 950 955
960Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val
965
970 975Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val
Gln 980 985 990Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln
Thr Tyr Val 995 1000 1005Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile
Arg Ala Ser Ala Asn 1010 1015 1020Leu Ala Ala Thr Lys Met Ser Glu
Cys Val Leu Gly Gln Ser Lys 1025 1030 1035Arg Val Asp Phe Cys Gly
Lys Gly Tyr His Leu Met Ser Phe Pro 1040 1045 1050Gln Ser Ala Pro
His Gly Val Val Phe Leu His Val Thr Tyr Val 1055 1060 1065Pro Ala
Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His 1070 1075
1080Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn
1085 1090 1095Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu
Pro Gln 1100 1105 1110Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly
Asn Cys Asp Val 1115 1120 1125Val Ile Gly Ile Val Asn Asn Thr Val
Tyr Asp Pro Leu Gln Pro 1130 1135 1140Glu Leu Asp Ser Phe Lys Glu
Glu Leu Asp Lys Tyr Phe Lys Asn 1145 1150 1155His Thr Ser Pro Asp
Val Asp Leu Gly Asp Ile Ser Gly Ile Asn 1160 1165 1170Ala Ser Val
Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu 1175 1180 1185Val
Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu 1190 1195
1200Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu
1205 1210 1215Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr
Ile Met 1220 1225 1230Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu
Lys Gly Cys Cys 1235 1240 1245Ser Cys Gly Ser Cys Cys Lys Phe Asp
Glu Asp Asp Ser Glu Pro 1250 1255 1260Val Leu Lys Gly Val Lys Leu
His Tyr Thr Trp Ser His Pro Gln 1265 1270 1275Phe Glu Lys
12801353846DNAArtificial SequenceNucleic acid sequence of full
length S protein of SARS-CoV-2 135atgttcgtgt tcctggtgct gctgcctctg
gtgagcagcc agtgcgtgaa tctgaccacc 60agaacccagc tgcctcctgc ctacaccaat
agcttcacca gaggagttta ttatcccgat 120aaggtgttca gaagtagtgt
attacatagt acccaggacc tgttcctacc tttcttcagt 180aacgtgacct
ggttccacgc catccacgtg agcggcacca atggcaccaa gagattcgac
240aatcctgtgc tgcctttcaa tgacggcgtg tacttcgcca gcaccgagaa
gagcaatatc 300atcagaggct ggatcttcgg caccaccttg gattccaaga
ctcagagcct gctgattgta 360aacaacgcta caaatgtggt gatcaaggtg
tgcgagttcc agttctgcaa tgaccctttc 420ctgggtgttt attatcataa
gaacaacaag agctggatgg agagcgagtt ccgcgtatat 480tcgtcggcta
ataattgcac cttcgagtac gtgagccagc ctttcctgat ggacctggag
540ggcaagcagg gcaatttcaa gaatctgaga gagttcgtgt tcaagaatat
cgacggctac 600ttcaagatct acagcaagca cacacccatt aatctggtga
gagacctgcc tcagggcttc 660agcgccctgg agcctctggt ggacctgcct
atcggcatca atatcaccag attccagacc 720ctgctggccc tgcacagatc
atatcttaca ccaggcgatt cgtcaagcgg ttggaccgct 780ggagctgcgg
catattacgt gggctacctg cagcctagaa ccttcctgct gaagtacaat
840gagaatggta cgataaccga cgcagttgat tgtgccctgg accctctgag
cgagaccaag 900tgcaccctga agagcttcac cgtggagaag ggcatctacc
agaccagcaa tttcagagtg 960cagcctaccg agagcatcgt gagattccct
aatatcacca atctgtgccc tttcggcgag 1020gtgttcaatg ccaccagatt
cgccagcgtg tacgcatgga accgcaagcg gataagcaat 1080tgcgtggccg
actacagcgt gctgtacaat agcgccagct tcagcacctt caaatgttat
1140ggtgtttcgc caacaaagct gaatgacctg tgcttcacca atgtgtacgc
cgacagcttc 1200gtgatcagag gcgacgaggt gagacagatc gcgccagggc
agaccggcaa gatcgccgac 1260tacaattaca agctgcctga cgacttcacc
ggctgcgtga tcgcgtggaa ctctaacaat 1320ctagattcga aagttggagg
caattacaat tacctgtaca gactgttcag aaagagcaat 1380ctgaagcctt
tcgagagaga catcagcacc gagatctacc aggccggcag cacaccgtgt
1440aatggcgtgg agggcttcaa ttgctacttc cctctgcaga gctacggctt
ccagcctacc 1500aatggcgtgg gctaccagcc ttacagagtg gtggtgctga
gcttcgagct gctgcacgct 1560cccgctaccg tgtgcggccc taagaagagc
accaatctgg tgaagaataa gtgcgtgaat 1620ttcaatttca atggtctaac
tggaacgggc gtgctgaccg agagcaataa gaagtttctt 1680ccctttcaac
aattcggcag agacatcgcc gacaccacag atgctgtaag agaccctcag
1740accctggaga tcctggacat cactccgtgt agcttcggcg gcgtgagcgt
gatcacaccg 1800ggtaccaata ccagcaatca ggtggccgtg ctgtaccagg
acgtgaattg caccgaggtg 1860cctgtggcca tccacgccga ccagctgact
cccacttgga gggtatattc cacgggaagc 1920aatgtgttcc agaccagagc
cggctgcctg atcggcgccg agcacgtgaa taatagctac 1980gagtgcgaca
tccctatcgg cgccggcatc tgcgccagct accagaccca gaccaatagc
2040cctagaagag ccagaagcgt ggccagccag agcatcatcg cctacaccat
gagcctgggc 2100gccgagaata gcgtggccta cagcaataat agcatcgcca
tccctaccaa tttcaccatc 2160agcgtgacca ccgaaatatt accagtctcc
atgaccaaga ccagcgtgga ctgcaccatg 2220tacatctgcg gcgacagcac
cgagtgcagc aatctgctgc tgcagtacgg cagcttctgc 2280acccagctga
atagagccct gaccggcatc gccgtggagc aggacaagaa tacccaggag
2340gtgttcgccc aggtgaagca gatctacaag actccgccga tcaaggactt
cggcggcttc 2400aatttcagcc aaatactccc agatccaagc aagcctagca
agaggagctt catcgaggac 2460ctgctgttca ataaggtgac cctggccgac
gccggcttca tcaagcagta cggcgactgc 2520ctaggtgata ttgcggcaag
agacctgatc tgcgcccaga agtttaacgg tttgacagta 2580ctacctcctc
tgctgaccga cgagatgata gcacaatata cgtcggcatt gctcgctggc
2640acgatcacat cgggctggac tttcggcgcc ggagcagcgt tgcaaatccc
tttcgccatg 2700cagatggcct acagattcaa tggcatcggc gtgacccaga
atgtgctgta cgagaatcag 2760aagctgatcg ccaatcagtt caatagcgcc
atcggcaaga tccaggacag cctgagcagc 2820accgccagcg ccctgggcaa
gctgcaggac gtggtgaatc agaatgccca ggccctgaat 2880accctggtga
agcagctgag cagcaatttc ggcgccatca gtagtgtact caacgatatc
2940ctgagcagac tggacaaggt ggaggccgag gtgcaaattg atcgtcttat
tactggcaga 3000ctgcagagcc tgcagaccta cgtgacccag cagctgatca
gagccgccga gatcagagcc 3060agcgccaatc tggccgccac caagatgagc
gagtgcgtgc tgggccagag caagagagtg 3120gacttctgcg gcaagggcta
ccacctgatg agcttccctc agagcgctcc acatggcgtg 3180gtgttcctgc
acgtgaccta cgtgcctgcc caggagaaga atttcaccac cgcacccgca
3240atctgccacg acggcaaggc ccacttccct agagagggcg tgttcgtgag
caatggcacc 3300cactggttcg tgacccagag aaatttctac gagcctcaga
tcatcaccac cgacaatacc 3360ttcgtgagcg gcaattgcga cgtggtgatc
gggatagtca ataatactgt ctacgaccct 3420ctgcagcctg agctggacag
cttcaaggag gagctggaca agtacttcaa gaatcacacc 3480agccctgacg
tggacctcgg tgatatttcg ggaatcaatg ccagcgtggt gaatatccag
3540aaggaaattg atcggctcaa cgaagtggcc aagaatctga atgagagcct
gatcgacctg 3600caggagctgg gcaagtacga gcagtacatc aagtggcctt
ggtacatctg gctgggcttc 3660atcgccggcc tgatcgccat cgtgatggtg
accatcatgc tgtgctgcat gacctcctgt 3720tgttcctgtt tgaaagggtg
ttgttcgtgt gggtcctgct gcaagttcga cgaggacgac 3780agcgagcctg
tgctgaaggg cgtgaagctg cactacacct ggagccaccc tcagttcgag 3840aagtga
38461368PRTHomo sapiensP2B-1G5 HCDR1(1)..(8) 136Gly Tyr Thr Phe Thr
Thr Tyr Val1 51378PRTHomo sapiensP2B-1G5 HCDR2(1)..(8) 137Ile Asn
Thr Asn Thr Gly Asn Pro1 513812PRTHomo sapiensP2B-1G5
HCDR3(1)..(12) 138Ser Cys Glu Ile Thr Thr Leu Gly Gly Met Asp Val1
5 101396PRTHomo sapiensP2B-1G5 LCDR1(1)..(6) 139Asn Ile Gly Ser Lys
Ser1 51404PRTHomo sapiensP2B-1G5
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 140Tyr Asp Ser
Xaa114111PRTHomo sapiensP2B-1G5 LCDR3(1)..(11) 141Gln Val Trp Asp
Ser Ile Ser Asp His Arg Val1 5 10142119PRTHomo sapiensP2B-1G5
VH(1)..(119) 142Gln Val Gln Leu Val Gln Ser Gly Ser Glu Leu Lys Lys
Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Thr Tyr 20 25 30Val Met Asn Trp Val Arg Gln Ala Pro Gly Gln
Gly Leu Glu Trp Met 35 40 45Gly Trp Ile Asn Thr Asn Thr Gly Asn Pro
Thr Tyr Ala Gln Gly Phe 50 55 60Thr Gly Arg Phe Val Phe Ser Leu Asp
Thr Ser Val Ser Thr Ala Ser65 70 75 80Leu Gln Ile Ser Ser Leu Lys
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ser Cys Glu Ile Thr Thr
Leu Gly Gly Met Asp Val Trp Gly Gln Gly 100 105 110Thr Thr Val Thr
Val Ser Ser 115143108PRTHomo sapiensP2B-1G5 VL(1)..(108) 143Ser Tyr
Val Leu Thr Gln Pro Pro Ser Val Ser Val Ala Pro Gly Lys1 5 10 15Thr
Ala Arg Ile Thr Cys Gly Gly Asn Asn Ile Gly Ser Lys Ser Val 20 25
30His Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr
35 40 45Tyr Asp Ser Asp Arg Pro Ser Gly Ile Pro Glu Arg Phe Ser Gly
Ser 50 55 60Asn Ser Gly Asn Thr Ala Thr Leu Thr Ile Ser Gly Val Glu
Ala Gly65 70 75 80Asp Glu Ala Asp Tyr Tyr Cys Gln Val Trp Asp Ser
Ile Ser Asp His 85 90 95Arg Val Phe Gly Gly Gly Thr Lys Leu Thr Val
Leu 100 105144357DNAHomo sapiensP2B-1G5 VHnu(1)..(357)
144caggtgcagc tggtgcaatc tgggtctgag ttgaagaagc ctggggcctc
agtgaaggtt 60tcctgcaagg cttctggata caccttcact acctatgtta tgaattgggt
gcgacaggcc 120cctggacaag ggcttgagtg gatgggatgg atcaacacca
acactgggaa cccaacgtat 180gcccagggct tcacaggacg gtttgtcttc
tccttggaca cctctgtcag cacggcatct 240ctgcagatca gcagcctaaa
ggctgaggac actgccgtgt attactgttc gtgtgaaata 300accaccttgg
gcggtatgga cgtctggggc caagggacca cggtcaccgt ctcctca
357145324DNAHomo sapiensP2B-1G5 VLnu(1)..(324) 145tcctatgtgc
tgactcagcc accctcagtg tcagtggccc caggaaagac ggccaggatt 60acctgtgggg
gaaacaacat tggaagtaaa agtgtgcact ggtaccagca gaagccaggc
120caggcccctg tgctggtcat ctattatgat agcgaccggc cctcagggat
ccctgagcga 180ttctctggct ccaactctgg gaacacggcc accctgacca
tcagcggggt cgaagccggg 240gatgaggccg actattactg tcaggtgtgg
gatagtatta gtgatcatcg ggtgttcggc 300ggagggacca agctgaccgt ccta
3241468PRTHomo sapiensP2B-1A1 HCDR1(1)..(8) 146Gly Gly Ser Ile Ser
Ser Tyr Tyr1 51477PRTHomo sapiensP2B-1A1 HCDR2(1)..(7) 147Ile Tyr
Tyr Ser Gly Ser Thr1 514814PRTHomo sapiensP2B-1A1 HCDR3(1)..(14)
148Ala Arg Leu Glu Arg Asp Trp Pro Leu Asp Ala Phe Asp Ile1 5
101499PRTHomo sapiensP2B-1A1 LCDR1(1)..(9) 149Ser Ser Asp Val Gly
Gly Tyr Asn Tyr1 51504PRTHomo sapiensP2B-1A1
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 150Asp Val Ser
Xaa115110PRTHomo sapiensP2B-1A1 LCDR3(1)..(10) 151Ser Ser Tyr Thr
Ser Asn Asn Thr Phe Ala1 5 10152120PRTHomo sapiensP2B-1A1
VH(1)..(120) 152Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys
Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser
Ile Ser Ser Tyr 20 25 30Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys
Gly Leu Glu Trp Ile 35 40 45Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn
Tyr Asn Pro Ser Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Val Asp Thr
Ser Lys Lys Gln Phe Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala
Ala Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Leu Glu Arg Asp Trp
Pro Leu Asp Ala Phe Asp Ile Trp Gly Gln 100 105 110Gly Thr Met Val
Thr Val Ser Ser 115 120153110PRTHomo sapiensP2B-1A1 VL(1)..(110)
153Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln1
5 10 15Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly
Tyr 20 25 30Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro
Lys Phe 35 40 45Met Ile Tyr Asp Val Ser Lys Arg Pro Ser Gly Val Ser
Asn Arg Phe 50 55 60Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr
Ile Ser Gly Leu65 70 75 80Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys
Ser Ser Tyr Thr Ser Asn 85 90 95Asn Thr Phe Ala Phe Gly Gly Gly Thr
Lys Leu Thr Val Leu 100 105 110154360DNAHomo sapiensP2B-1A1
VHnu(1)..(360) 154caggtgcagc tgcaggagtc gggcccagga ctggtgaagc
cttcggagac cctgtccctc 60acctgcactg tctctggtgg ctccatcagt agttactact
ggagctggat ccggcagccc 120ccagggaagg gactggagtg gattgggtat
atctattaca gtgggagcac caactacaac 180ccctccctca agagtcgagt
caccatatca gtagacacgt ccaagaagca gttctccctg 240aagctgagct
ctgtgaccgc tgcggacacg gccgtgtatt actgtgcgag gctcgaacga
300gactggccac ttgatgcttt tgatatctgg ggccaaggga caatggtcac
cgtctcctca 360155330DNAHomo sapiensP2B-1A1 VLnu(1)..(330)
155cagtctgccc tgactcagcc tgcctccgtg tctgggtctc ctggacagtc
gatcaccatc 60tcctgcactg gaaccagcag tgacgttggt ggttataact atgtctcctg
gtaccaacag 120cacccaggca aagcccccaa attcatgatt tatgatgtca
gtaagcggcc ctcaggggtt 180tctaatcgct tctctggctc caagtctggc
aacacggcct ccctgaccat ctctgggctc 240caggctgagg acgaggctga
ttattactgc agctcatata caagcaacaa cactttcgcg 300ttcggcggag
ggaccaagct gaccgtccta 3301568PRTHomo sapiensP2C-1D7 HCDR1(1)..(8)
156Gly Phe Thr Val Ser Ser Asn Tyr1 51577PRTHomo sapiensP2C-1D7
HCDR2(1)..(7) 157Ile Tyr Ser Gly Gly Ser Thr1 515812PRTHomo
sapiensP2C-1D7 HCDR3(1)..(12) 158Ala Arg Glu Leu Tyr Glu Val Gly
Ala Thr Asp Tyr1 5 1015911PRTHomo sapiensP2C-1D7 LCDR1(1)..(11)
159Gln Ser Leu Val Tyr Ser Asp Gly Asn Thr Tyr1 5 101604PRTHomo
sapiensP2C-1D7 LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent.
160Lys Val Ser Xaa11619PRTHomo sapiensP2C-1D7 LCDR3(1)..(9) 161Met
Gln Arg Tyr Thr Leu Ala Gly Val1 5162118PRTHomo sapiensP2C-1D7
VH(1)..(118) 162Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln
Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Val Ser Ser Asn 20 25 30Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr
Tyr Ala Asp Ser Val Lys 50 55 60Gly Arg Phe Thr Ile Ser Arg Asp Asn
Ser Lys Asn Thr Leu Tyr Leu65 70 75 80Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Glu Leu Tyr Glu Val
Gly Ala Thr Asp Tyr Trp Gly Gln Gly Thr 100 105 110Leu Val Thr Val
Ser Ser 115163112PRTHomo sapiensP2C-1D7 VL(1)..(112) 163Asp Val Val
Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly1 5 10 15Gln Pro
Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val Tyr Ser 20 25 30Asp
Gly Asn Thr Tyr Leu Asn Trp Phe Gln Gln Arg Pro Gly Gln Ser 35 40
45Pro Arg Arg Leu Ile Tyr Lys Val Ser Asn Trp Asp Ser Gly Val Pro
50 55 60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys
Ile65 70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys
Met Gln Arg 85 90 95Tyr Thr Leu Ala Gly Val Phe Gly Pro Gly Thr Lys
Val Asp Ile Lys 100 105 110164354DNAHomo sapiensP2C-1D7
VHnu(1)..(354) 164gaggtgcagc tggtggagtc tggaggaggc ttgatccagc
ctggggggtc cctgagactc 60tcctgtgcag cctctgggtt caccgtcagt agcaactaca
tgagctgggt ccgccaggct 120ccagggaagg ggctggagtg ggtctcagtt
atttatagcg gtggtagcac atactacgca 180gactccgtga agggccgatt
caccatctcc agagacaatt ccaagaacac gctgtatctt 240caaatgaaca
gcctgagagc cgaggacacg gccgtgtatt actgtgcgag agaattgtac
300gaagtgggag ctacggacta ctggggccag ggaaccctgg tcaccgtctc ctca
354165336DNAHomo sapiensP2C-1D7 VLnu(1)..(336) 165gatgttgtga
tgactcagtc tccactctcc ctgcccgtca cccttggaca gccggcctcc 60atctcctgca
ggtctagtca aagcctcgta tacagtgatg gaaacaccta cttgaattgg
120tttcagcaga ggccaggcca atctccaagg cgcctaattt ataaggtttc
taactgggac 180tctggggtcc cagacagatt cagcggcagt gggtcaggca
ctgatttcac actgaaaatc 240agcagggtgg aggctgagga tgttggggtt
tattactgca tgcaacggta cacactggcc 300ggcgttttcg gccctgggac
caaagtggat atcaaa 3361668PRTHomo sapiensP2B-1A10 HCDR1(1)..(8)
166Gly Phe Thr Val Ser Ser Asn Tyr1 51677PRTHomo sapiensP2B-1A10
HCDR2(1)..(7) 167Ile Tyr Ser Gly Gly Ser Thr1 516815PRTHomo
sapiensP2B-1A10 HCDR3(1)..(15) 168Ala Arg Glu Gly Pro Lys Ser Ile
Thr Gly Thr Ala Phe Asp Ile1 5 10 151696PRTHomo sapiensP2B-1A10
LCDR1(1)..(6) 169Gln Asp Ile Ser Asn Tyr1 51704PRTHomo
sapiensP2B-1A10 LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent.
170Asp Ala Ser Xaa117110PRTHomo sapiensP2B-1A10 LCDR3(1)..(10)
171Gln Gln Tyr Asp Asn Leu Pro Met Tyr Thr1 5 10172121PRTHomo
sapiensP2B-1A10 VH(1)..(121) 172Glu Val Gln Leu Val Glu Ser Gly Gly
Gly Leu Ile Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Val Ser Ser Asn
20 25 30Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser
Val Lys 50 55 60Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr Leu65 70 75 80Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr Cys Ala 85 90 95Arg Glu Gly Pro Lys Ser Ile Thr Gly Thr
Ala Phe Asp Ile Trp Gly 100 105 110Gln Gly Thr Ile Val Thr Val Ser
Ser 115 120173108PRTHomo sapiensP2B-1A10 VL(1)..(108) 173Asp Ile
Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp
Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp Ile Ser Asn Tyr 20 25
30Phe Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
35 40 45Tyr Asp Ala Ser Asn Leu Glu Thr Gly Val Pro Ser Arg Phe Ser
Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser Leu
Gln Pro65 70 75 80Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Tyr Asp
Asn Leu Pro Met 85 90 95Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile
Lys 100 105174363DNAHomo sapiensP2B-1A10 VHnu(1)..(363)
174gaggtgcagc tggtggagtc tggaggaggc ttgatccagc ctggggggtc
cctgagactc 60tcctgtgcag cctctgggtt caccgtcagt agcaactaca tgagctgggt
ccgccaggct 120ccagggaagg ggctggagtg ggtctcagtt atttatagcg
gtggtagcac atactacgca 180gactccgtga agggccgatt caccatctcc
agagacaatt ccaagaacac gctgtatctt 240caaatgaaca gcctgagagc
cgaggacacg gccgtttatt actgtgcgag agagggccca 300aagtctatta
cagggacggc ttttgatatc tggggccaag ggacaattgt caccgtctcc 360tca
363175324DNAHomo sapiensP2B-1A10 VLnu(1)..(324) 175gacatccaga
tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc
aggcgagtca ggacattagc aactatttta attggtatca gcagaaacca
120gggaaagccc ctaagctcct gatctacgat gcatccaatt tggaaacagg
ggtcccatca 180aggttcagtg gaagtggatc tgggacagat tttactttca
ccatcagcag cctgcagcct 240gaagatattg caacatatta ctgtcaacag
tatgataatc tccccatgta cacttttggc 300caggggacca agctggagat caaa
32417610PRTHomo sapiensP2B-1D9 HCDR1(1)..(10) 176Gly Phe Ser Leu
Ser Thr Ser Gly Val Gly1 5 101777PRTHomo sapiensP2B-1D9
HCDR2(1)..(7) 177Ile Tyr Trp Asp Asp Asp Lys1 517816PRTHomo
sapiensP2B-1D9 HCDR3(1)..(16) 178Ala His Thr Arg Ile Leu Tyr Tyr
Gly Ser Gly Ser Tyr Tyr Asp Tyr1 5 10 151798PRTHomo sapiensP2B-1D9
LCDR1(1)..(8) 179Ser Ser Asn Ile Gly Ser Asn Tyr1 51804PRTHomo
sapiensP2B-1D9 LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent.
180Ser Asn Asn Xaa118111PRTHomo sapiensP2B-1D9 LCDR3(1)..(11)
181Ala Ala Trp Asp Asp Ser Leu Ser Gly Val Val1 5 10182124PRTHomo
sapiensP2B-1D9 VH(1)..(124) 182Gln Ile Thr Leu Lys Glu Ser Gly Pro
Thr Leu Val Lys Pro Thr Gln1 5 10 15Thr Leu Thr Leu Thr Cys Thr Phe
Ser Gly Phe Ser Leu Ser Thr Ser 20 25 30Gly Val Gly Val Gly Trp Ile
Arg Gln Pro Pro Gly Lys Ala Leu Glu 35 40 45Trp Leu Ala Leu Ile Tyr
Trp Asp Asp Asp Lys Tyr Tyr Ser Pro Ser 50 55 60Leu Lys Ser Arg Leu
Thr Ile Thr Lys Asp Thr Ser Lys Asn Gln Val65 70 75 80Val Leu Thr
Met Thr Asn Met Asp Pro Val Asp Thr Ala Thr Tyr Tyr 85 90 95Cys Ala
His Thr Arg Ile Leu Tyr Tyr Gly Ser Gly Ser Tyr Tyr Asp 100 105
110Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115
120183110PRTHomo sapiensP2B-1D9 VL(1)..(110) 183Gln Ser Val Leu Thr
Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile
Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Tyr Val Tyr
Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr
Ser Asn Asn Gln Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly
Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Arg65 70 75
80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu
85 90 95Ser Gly Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100
105 110184372DNAHomo sapiensP2B-1D9 VHnu(1)..(372) 184cagatcacct
tgaaggagtc tggtcctacg ctggtgaaac ccacacagac cctcacgctg 60acctgcacct
tctctgggtt ctcactcagc actagtggag tgggtgtggg ctggatccgt
120cagcccccag gaaaggccct ggagtggctt gcactcattt attgggatga
tgataaatac 180tacagcccat ctctgaagag caggctcacc atcaccaagg
acacctccaa aaaccaggtg 240gtccttacaa tgaccaacat ggaccctgtg
gacacagcca catattactg tgcacacact 300cgcatcttat actatggttc
ggggagttat tatgactact ggggccaggg aaccctggtc 360accgtctcct ca
372185330DNAHomo sapiensP2B-1D9 VLnu(1)..(330) 185cagtctgtgc
tgactcagcc accctcagcg tctgggaccc ccgggcagag ggtcaccatc 60tcttgttctg
gaagcagctc caacatcgga agtaattatg tatactggta ccagcagctc
120ccaggaacgg cccccaaact cctcatctat agtaataatc agcggccctc
aggggtccct 180gaccgattct ctggctccaa gtctggcacc tcagcctccc
tggccatcag tgggctccgg 240tccgaggatg aggctgatta ttactgtgca
gcatgggatg acagcctgag tggtgtggta 300ttcggcggag ggaccaagct
gaccgtccta 33018610PRTHomo sapiensP2B-1E4 HCDR1(1)..(10) 186Gly Phe
Ser Leu Ser Thr Ser Gly Val Gly1 5 101877PRTHomo sapiensP2B-1E4
HCDR2(1)..(7) 187Ile Tyr Trp Asp Asp Asp Lys1 518811PRTHomo
sapiensP2B-1E4 HCDR3(1)..(11) 188Ala His Gln Ile Val Ala Thr Ile
Ile Asp Tyr1 5 101899PRTHomo sapiensP2B-1E4 LCDR1(1)..(9) 189Ser
Ser Asp Val Gly Gly Tyr Asn Tyr1 51904PRTHomo sapiensP2B-1E4
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 190Asp Val Ser
Xaa11919PRTHomo sapiensP2B-1E4 LCDR3(1)..(9) 191Ser Ser Tyr Thr Ser
Ser Ser Val Val1 5192119PRTHomo sapiensP2B-1E4 VH(1)..(119) 192Gln
Ile Thr Leu Lys Glu Ser Gly Pro Thr Leu Val Lys Pro Thr Gln1 5 10
15Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser Leu Ser Thr Ser
20 25 30Gly Val Gly Val Gly Trp Ile Arg Gln Pro Pro Gly Lys Ala Leu
Glu 35 40 45Trp Leu Ala Leu Ile Tyr Trp Asp Asp Asp Lys Arg Tyr Ser
Pro Ser 50 55 60Leu Lys Ser Arg Leu Thr Ile Thr Lys Asp Thr Ser Lys
Asn Gln Val65 70 75 80Val Leu Thr Met Thr Asn Met Asp Pro Val Asp
Thr Ala Thr Tyr Tyr 85 90 95Cys Ala His Gln Ile Val Ala Thr Ile Ile
Asp Tyr Trp Gly Gln Gly 100 105 110Thr Leu Val Thr Val Ser Ser
115193109PRTHomo sapiensP2B-1E4 VL(1)..(109) 193Gln Ser Ala Leu Thr
Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln1 5 10 15Ser Ile Thr Ile
Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn Tyr Val
Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45Met Ile
Tyr Asp Val Ser Lys Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55 60Ser
Gly Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65 70 75
80Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser
85 90 95Ser Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100
105194357DNAHomo sapiensP2B-1E4 VHnu(1)..(357) 194cagatcacct
tgaaggagtc tggtcctacg ctggtgaaac ccacacagac cctcacgctg 60acctgcacct
tctctgggtt ctcactcagc actagtggag tgggtgtggg ctggatccgt
120cagcccccag gaaaggccct ggagtggctt gcactcattt attgggatga
tgataagcgc 180tacagcccat ctctgaagag caggctcacc atcaccaagg
acacctccaa aaaccaggtg 240gtccttacaa tgaccaacat ggaccctgtg
gacacagcca catattactg tgcacaccaa 300atagtggcta cgattattga
ctactggggc cagggaaccc tggtcaccgt ctcctca 357195327DNAHomo
sapiensP2B-1E4 VLnu(1)..(327) 195cagtctgccc tgactcagcc tgcctccgtg
tctgggtctc ctggacagtc gatcaccatc 60tcctgcactg gaaccagcag tgacgttggt
ggttataact atgtctcctg gtaccaacag 120cacccaggca aagcccccaa
actcatgatt tatgatgtca gtaagcggcc ctcaggggtt 180tctaatcgct
tctctggctc caagtctggc aacacggcct ccctgaccat ctctgggctc
240caggctgagg acgaggctga ttattactgc agctcatata caagcagcag
cgtggtattc 300ggcggaggga ccaagctgac cgtccta 3271968PRTHomo
sapiensP2B-1G1 HCDR1(1)..(8) 196Gly Phe Thr Val Ser Ser Asn Tyr1
51977PRTHomo sapiensP2B-1G1 HCDR2(1)..(7) 197Ile Tyr Ser Gly Gly
Ser Thr1 519811PRTHomo sapiensP2B-1G1 HCDR3(1)..(11) 198Ala Arg Asp
Tyr Gly Asp Tyr Trp Phe Asp Pro1 5 101997PRTHomo sapiensP2B-1G1
LCDR1(1)..(7) 199Gln Ser Val Ser Ser Ser Tyr1 52004PRTHomo
sapiensP2B-1G1 LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent.
200Gly Ala Ser Xaa12019PRTHomo sapiensP2B-1G1 LCDR3(1)..(9) 201Gln
Gln Tyr Gly Ser Ser Pro Arg Thr1 5202117PRTHomo sapiensP2B-1G1
VH(1)..(117) 202Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln
Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Val Ser Ser Asn 20 25 30Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr
Tyr Ala Asp Ser Val Lys 50 55 60Gly Arg Phe Thr Ile Ser Arg Asp Asn
Ser Lys Asn Thr Leu Tyr Leu65 70 75 80Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Asp Tyr Gly Asp Tyr
Trp Phe Asp Pro Trp Gly Gln Gly Thr Leu 100 105 110Val Thr Val Ser
Ser 115203108PRTHomo sapiensP2B-1G1 VL(1)..(108) 203Glu Ile Val Leu
Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala
Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Tyr Leu
Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile
Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55
60Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65
70 75 80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser
Pro 85 90 95Arg Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100
105204351DNAHomo sapiensP2B-1G1 VHnu(1)..(351) 204gaggtgcagc
tggtggagtc tgggggaggc ttggtccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctggatt caccgtcagt agcaactaca tgagctgggt ccgccaggct
120ccagggaagg ggctggagtg ggtctcagtt atttatagcg gtggtagcac
atactacgca 180gactccgtga agggcagatt caccatctcc agagacaatt
ccaagaacac gctgtatctt 240caaatgaaca gcctgagagc cgaggacacg
gctgtgtatt actgtgcgag agactacggt 300gactactggt tcgacccctg
gggccaggga accctggtca ccgtctcctc a 351205324DNAHomo sapiensP2B-1G1
VLnu(1)..(324) 205gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt
ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagc agcagctact
tagcctggta ccagcagaaa 120cctggccagg ctcccaggct cctcatctat
ggtgcatcca gcagggccac tggcatccca 180gacaggttca gtggcagtgg
gtctgggaca gacttcactc tcaccatcag cagactggag 240cctgaagatt
ttgcagtgta ttactgtcag cagtatggta gctcaccgag gacttttggc
300caggggacca agctggagat caaa 3242068PRTHomo sapiensP4A-2D9
HCDR1(1)..(8) 206Gly Phe Thr Phe Ser Ser Tyr Gly1 52078PRTHomo
sapiensP4A-2D9 HCDR2(1)..(8) 207Ile Ser Asp Asp Gly Ser Asn Gln1
520821PRTHomo sapiensP4A-2D9 HCDR3(1)..(21) 208Ala Lys Arg Gly Gly
Tyr Cys Ser Thr Thr Ser Cys Leu Val Arg Trp1 5 10 15Val Tyr Phe Asp
Tyr 202096PRTHomo sapiensP4A-2D9 LCDR1(1)..(6) 209Gln Phe Ile Ser
Ser Tyr1 52104PRTHomo sapiensP4A-2D9
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 210Ala Thr Ser
Xaa12118PRTHomo sapiensP4A-2D9 LCDR3(1)..(8) 211Gln Gln Ser Tyr Asn
Thr Leu Thr1 5212128PRTHomo sapiensP4A-2D9 VH(1)..(128) 212Gln Val
Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10 15Ser
Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25
30Gly Met His Trp Val Arg Gln Ser Pro Gly Lys Gly Leu Glu Trp Val
35 40 45Ala Val Ile Ser Asp Asp Gly Ser Asn Gln Tyr Tyr Ala Asp Ser
Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr65 70 75 80Leu Glu Ile Asn Ser Leu Arg Val Glu Asp Thr Ala
Val Tyr Tyr Cys 85 90 95Ala Lys Arg Gly Gly Tyr Cys Ser Thr Thr Ser
Cys Leu Val Arg Trp 100 105 110Val Tyr Phe Asp Tyr Trp Gly Gln Gly
Thr Leu Val Thr Val Ser Ser 115 120 125213106PRTHomo sapiensP4A-2D9
VL(1)..(106) 213Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala
Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Phe
Ile Ser Ser Tyr 20 25 30Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala
Pro Lys Leu Leu Ile 35 40 45Tyr Ala Thr Ser Ile Leu Gln Thr Gly Val
Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Leu
Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe Ala Thr Tyr Tyr
Cys Gln Gln Ser Tyr Asn Thr Leu Thr 85 90 95Phe Gly Pro Gly Thr Lys
Val Asp Ile Lys 100 105214384DNAHomo sapiensP4A-2D9 VHnu(1)..(384)
214caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc
cctgagactc 60tcctgtgcag cctctggatt caccttcagt agctatggca tgcactgggt
ccgccagtct 120ccaggcaagg ggctggagtg ggtggcagtt atatcagatg
atggaagtaa tcaatactat 180gcagactccg tgaagggccg attcaccatc
tccagagaca attccaagaa cacgctgtat 240ctggaaatca acagcctgag
agttgaggac acggctgtgt attactgtgc gaaaaggggc 300ggatattgta
gtactaccag ctgcctcgtt aggtgggtct actttgacta ctggggccag
360ggaaccctgg tcaccgtctc ctca 384215318DNAHomo sapiensP4A-2D9
VLnu(1)..(318) 215gacatccaga tgacccagtc tccatcctcc ctgtctgcat
ctgtaggaga cagagtcacc 60atcacttgcc gggcaagtca gttcattagc agctacttaa
attggtatca gcagaaacca 120gggaaagccc ctaagctcct gatctatgct
acatccattt tgcaaactgg ggtcccatca 180aggttcagtg gcagtggatc
tgggacagat ttcactctca ccatcagcag tctgcaacct 240gaagattttg
caacttacta ctgtcaacag agttacaata cccttacttt cggccctggg
300accaaagtcg atatcaaa 31821610PRTHomo sapiensP5A-2G7
HCDR1(1)..(10) 216Gly Asp Ser Val Ser Ser Gly Ser Tyr Tyr1 5
102177PRTHomo sapiensP5A-2G7 HCDR2(1)..(7) 217Ile Tyr Tyr Ser Gly
Ser Thr1 521820PRTHomo sapiensP5A-2G7 HCDR3(1)..(20) 218Ala Arg Glu
Arg Cys Tyr Tyr Gly Ser Gly Arg Ala Pro Arg Cys Val1 5 10 15Trp Phe
Asp Pro 202199PRTHomo sapiensP5A-2G7 LCDR1(1)..(9) 219Ser Ser Asp
Val Gly Gly Tyr Asn Tyr1 52204PRTHomo sapiensP5A-2G7
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 220Asp Val Ser
Xaa122111PRTHomo sapiensP5A-2G7 LCDR3(1)..(11) 221Ser Ser Tyr Thr
Ser Ser Ser Thr Leu Val Val1 5 10222128PRTHomo sapiensP5A-2G7
VH(1)..(128) 222Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys
Pro Ser Glu1 5 10 15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser
Val Ser Ser Gly 20 25 30Ser Tyr Tyr Trp Ser Trp Ile Arg Gln Pro Pro
Gly Lys Gly Leu Glu 35 40 45Trp Ile Gly Tyr Ile Tyr Tyr Ser Gly Ser
Thr Asn Tyr Asn Pro Ser 50 55 60Leu Lys Ser Arg Val Thr Ile Ser Val
Asp Thr Ser Lys Asn Gln Phe65 70 75 80Ser Leu Lys Leu Ser Ser Val
Thr Ala Ala Asp Thr Ala Val Tyr Tyr 85 90 95Cys Ala Arg Glu Arg Cys
Tyr Tyr Gly Ser Gly Arg Ala Pro Arg Cys 100 105 110Val Trp Phe Asp
Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120
125223111PRTHomo sapiensP5A-2G7 VL(1)..(111) 223Gln Ser Ala Leu Thr
Gln Pro Ala Ser Val Ser Gly Ser Pro
Gly Gln1 5 10 15Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp Val
Gly Gly Tyr 20 25 30Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly Lys
Ala Pro Lys Leu 35 40 45Met Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly
Val Ser Asn Arg Phe 50 55 60Ser Gly Ser Lys Ser Gly Asn Thr Ala Ser
Leu Thr Ile Ser Gly Leu65 70 75 80Gln Ala Glu Asp Glu Ala Asp Tyr
Tyr Cys Ser Ser Tyr Thr Ser Ser 85 90 95Ser Thr Leu Val Val Phe Gly
Gly Gly Thr Lys Leu Thr Val Leu 100 105 110224384DNAHomo
sapiensP5A-2G7 VHnu(1)..(384) 224caggtgcagc tgcaggagtc gggcccagga
ctggtgaagc cttcggagac cctgtccctc 60acctgcactg tctctggtga ctccgtcagc
agtggtagtt actactggag ctggatccgg 120cagcccccag ggaagggact
ggagtggatt gggtatatct attacagtgg gagcaccaac 180tacaacccct
ccctcaagag tcgagtcacc atatcagtag acacgtccaa gaaccagttc
240tccctgaagc tgagctctgt gaccgctgcg gacacggccg tgtattactg
tgcgagagag 300cgatgttact atggttcagg gagagccccc cgttgtgtct
ggttcgaccc ctggggccag 360ggaaccctgg tcaccgtctc ctca
384225333DNAHomo sapiensP5A-2G7 VLnu(1)..(333) 225cagtctgccc
tgactcagcc tgcctccgtg tctgggtctc ctggacagtc gatcaccatc 60tcctgcactg
gaaccagcag tgacgttggt ggttataact atgtctcctg gtaccaacaa
120cacccaggca aagcccccaa actcatgatt tatgatgtca gtaatcggcc
ctcaggggtt 180tctaatcgct tctctggctc caagtctggc aacacggcct
ccctgaccat ctctgggctc 240caggctgagg acgaggctga ttattactgc
agctcatata caagcagcag cactctcgtg 300gtattcggcg gagggaccaa
gctgaccgtc cta 3332268PRTHomo sapiensP5A-3C8 HCDR1(1)..(8) 226Gly
Phe Thr Val Ser Ser Asn Tyr1 52277PRTHomo sapiensP5A-3C8
HCDR2(1)..(7) 227Ile Tyr Ser Gly Gly Ser Thr1 522811PRTHomo
sapiensP5A-3C8 HCDR3(1)..(11) 228Ala Arg Asp Leu Gln Glu His Gly
Met Asp Val1 5 102296PRTHomo sapiensP5A-3C8 LCDR1(1)..(6) 229Gln
Gly Ile Ser Ser Tyr1 52304PRTHomo sapiensP5A-3C8
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 230Ala Ala Ser
Xaa123111PRTHomo sapiensP5A-3C8 LCDR3(1)..(11) 231Gln His Leu Asn
Ser Tyr Pro Pro Gly Tyr Thr1 5 10232117PRTHomo sapiensP5A-3C8
VH(1)..(117) 232Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln
Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Val Ser Ser Asn 20 25 30Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ser Phe Ile Tyr Ser Gly Gly Ser Thr Tyr
Tyr Ala Asp Ser Val Lys 50 55 60Gly Arg Phe Thr Ile Ser Arg Asp Asn
Ser Lys Asn Thr Leu Tyr Leu65 70 75 80Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Asp Leu Gln Glu His
Gly Met Asp Val Trp Gly Gln Gly Thr Thr 100 105 110Val Thr Val Ser
Ser 115233109PRTHomo sapiensP5A-3C8 VL(1)..(109) 233Asp Ile Gln Leu
Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr 20 25 30Leu Ala
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr
Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln His Leu Asn Ser Tyr Pro
Pro 85 90 95Gly Tyr Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100
105234351DNAHomo sapiensP5A-3C8 VHnu(1)..(351) 234gaggtgcagc
tggtggagtc tggaggaggc ttgatccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctgggtt caccgtcagt agcaactaca tgagctgggt ccgccaggct
120ccagggaagg ggctggaatg ggtctcattt atttatagcg gtggtagtac
atactacgca 180gactccgtga agggccgatt caccatctcc agagacaatt
ccaagaacac gctgtatctt 240caaatgaaca gcctgagagc cgaggacacg
gccgtgtatt actgtgcgag agatctacag 300gaacacggta tggacgtctg
gggccaaggg accacggtca ccgtctcctc a 351235327DNAHomo sapiensP5A-3C8
VLnu(1)..(327) 235gacatccagt tgacccagtc tccatcctcc ctgtctgcat
ctgtaggaga cagagtcacc 60atcacttgcc gggccagtca gggcattagc agttatttag
cctggtatca gcaaaaacca 120gggaaagccc ctaagctcct gatctatgct
gcatccactt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc
tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg
caacttatta ctgtcaacac cttaatagtt accctccggg gtacactttt
300ggccagggga ccaagctgga gatcaaa 3272368PRTHomo sapiensP5A-1D2
HCDR1(1)..(8) 236Gly Phe Ile Val Ser Ser Asn Tyr1 52377PRTHomo
sapiensP5A-1D2 HCDR2(1)..(7) 237Ile Tyr Ser Gly Gly Ser Thr1
523815PRTHomo sapiensP5A-1D2 HCDR3(1)..(15) 238Ala Arg Ala Leu Gln
Val Gly Ala Thr Ser Asp Tyr Phe Asp Tyr1 5 10 152399PRTHomo
sapiensP5A-1D2 LCDR1(1)..(9) 239Ser Ser Asn Ile Gly Ala Gly Tyr
Asp1 52404PRTHomo sapiensP5A-1D2
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 240Gly Asn Ser
Xaa124111PRTHomo sapiensP5A-1D2 LCDR3(1)..(11) 241Gln Ser Cys Asp
Ser Ser Leu Ser Val Val Val1 5 10242121PRTHomo sapiensP5A-1D2
VH(1)..(121) 242Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln
Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ile
Val Ser Ser Asn 20 25 30Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ser Ile Ile Tyr Ser Gly Gly Ser Thr Tyr
Tyr Ala Asp Ser Val Lys 50 55 60Gly Arg Phe Thr Ile Ser Arg Asp Asn
Ser Asn Asn Thr Leu Tyr Leu65 70 75 80Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Ala Leu Gln Val Gly
Ala Thr Ser Asp Tyr Phe Asp Tyr Trp Gly 100 105 110Gln Gly Thr Leu
Val Thr Val Ser Ser 115 120243111PRTHomo sapiensP5A-1D2
VL(1)..(111) 243Gln Ser Val Leu Thr Gln Pro Pro Ser Val Ser Gly Ala
Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Thr Gly Ser Ser Ser Asn
Ile Gly Ala Gly 20 25 30Tyr Asp Val His Trp Tyr Gln Gln Leu Pro Gly
Thr Ala Pro Lys Leu 35 40 45Leu Ile Tyr Gly Asn Ser Asn Arg Pro Ser
Gly Val Pro Asp Arg Phe 50 55 60Ser Gly Ser Lys Ser Gly Thr Ser Ala
Ser Leu Ala Ile Thr Gly Leu65 70 75 80Gln Ala Glu Asp Glu Thr Asp
Tyr Tyr Cys Gln Ser Cys Asp Ser Ser 85 90 95Leu Ser Val Val Val Phe
Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 110244363DNAHomo
sapiensP5A-1D2 VHnu(1)..(363) 244gaggtgcagc tggtggagtc tggaggaggc
ttgatccagc ctggggggtc cctgagactc 60tcctgtgcag cctctgggtt catcgtcagt
agcaactaca tgagctgggt ccgccaggct 120ccagggaagg ggctggagtg
ggtctcaatt atttatagcg gtggtagcac atactacgca 180gactccgtga
agggccgatt caccatctcc agagacaatt ccaacaacac gctgtatctt
240caaatgaaca gcctgagagc cgaggacacg gccgtatatt actgtgcgag
agccctccag 300gtgggagcta cttcggacta ctttgactac tggggccagg
gaaccctggt caccgtctcc 360tca 363245333DNAHomo sapiensP5A-1D2
VLnu(1)..(333) 245cagtctgtgc tgacgcagcc gccctcagtg tctggggccc
cagggcagag ggtcaccatc 60tcctgcactg ggagcagctc caacatcggg gcaggttatg
atgtacactg gtaccagcaa 120cttccaggaa cagcccccaa actcctcatc
tatggtaaca gcaatcggcc ctcaggggtc 180cctgaccgat tctctggctc
caagtctggc acctcagcct ccctggccat cactgggctc 240caggctgaag
atgagactga ttattactgc cagtcctgtg acagcagcct gagtgttgtg
300gtattcggcg gagggaccaa gctgaccgtc cta 3332468PRTHomo
sapiensP5A-2F11 HCDR1(1)..(8) 246Gly Tyr Thr Phe Thr Ser Tyr Asp1
52478PRTHomo sapiensP5A-2F11 HCDR2(1)..(8) 247Met Asn Pro Asn Ser
Gly Asn Thr1 524815PRTHomo sapiensP5A-2F11 HCDR3(1)..(15) 248Ala
Arg Tyr Ile Val Val Val Pro Ala Ala Lys Gly Phe Asp Pro1 5 10
1524912PRTHomo sapiensP5A-2F11 LCDR1(1)..(12) 249Gln Ser Val Leu
Tyr Ser Ser Asn Asn Lys Asn Tyr1 5 102504PRTHomo sapiensP5A-2F11
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 250Trp Ala Ser
Xaa12519PRTHomo sapiensP5A-2F11 LCDR3(1)..(9) 251Gln Gln Tyr Tyr
Ser Thr Pro Leu Thr1 5252122PRTHomo sapiensP5A-2F11 VH(1)..(122)
252Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala1
5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser
Tyr 20 25 30Asp Ile Asn Trp Val Arg Gln Ala Thr Gly Gln Gly Leu Glu
Trp Met 35 40 45Gly Trp Met Asn Pro Asn Ser Gly Asn Thr Gly Tyr Ala
Gln Lys Phe 50 55 60Gln Gly Arg Val Thr Met Thr Arg Asn Thr Ser Ile
Ser Thr Ala Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg Ser Glu Asp
Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Tyr Ile Val Val Val Pro Ala
Ala Lys Gly Phe Asp Pro Trp 100 105 110Gly Gln Gly Thr Leu Val Thr
Val Ser Ser 115 120253113PRTHomo sapiensP5A-2F11 VL(1)..(113)
253Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly1
5 10 15Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr
Ser 20 25 30Ser Asn Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro
Gly Gln 35 40 45Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu
Ser Gly Val 50 55 60Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp
Phe Thr Leu Thr65 70 75 80Ile Ser Ser Leu Gln Ala Glu Asp Val Ala
Val Tyr Tyr Cys Gln Gln 85 90 95Tyr Tyr Ser Thr Pro Leu Thr Phe Gly
Gly Gly Thr Lys Val Glu Ile 100 105 110Lys254366DNAHomo
sapiensP5A-2F11 VHnu(1)..(366) 254caggtgcagc tggtgcagtc tggggctgag
gtgaagaagc ctggggcctc agtgaaggtc 60tcctgcaagg cttctggata caccttcacc
agttatgata tcaactgggt gcgacaggcc 120actggacaag ggcttgagtg
gatgggatgg atgaacccta acagtggtaa cacaggctat 180gcacagaagt
tccagggcag agtcaccatg accaggaaca cctccataag cacagcctac
240atggagctga gcagcctgag atctgaggac acggccgtgt attactgtgc
gagatatatt 300gtagtagtac cagctgcaaa agggttcgac ccctggggcc
agggaaccct ggtcaccgtc 360tcctca 366255339DNAHomo sapiensP5A-2F11
VLnu(1)..(339) 255gacatcgtga tgacccagtc tccagactcc ctggctgtgt
ctctgggcga gagggccacc 60atcaactgca agtccagcca gagtgtttta tacagctcca
acaataagaa ctacttagct 120tggtaccagc agaaaccagg acagcctcct
aagctgctca tttactgggc atctacccgg 180gaatccgggg tccctgaccg
attcagtggc agcgggtctg ggacagattt cactctcacc 240atcagcagcc
tgcaggctga agatgtggca gtttattact gtcagcaata ttatagtact
300cctctcactt tcggcggagg gaccaaggtg gagatcaaa 3392568PRTHomo
sapiensP5A-2E1 HCDR1(1)..(8) 256Gly Tyr Ser Phe Thr Ser Tyr Trp1
52578PRTHomo sapiensP5A-2E1 HCDR2(1)..(8) 257Ile Tyr Pro Gly Asp
Ser Asp Thr1 525812PRTHomo sapiensP5A-2E1 HCDR3(1)..(12) 258Ala Gln
Thr Ser Val Thr Arg Asn Trp Phe Asp Pro1 5 102596PRTHomo
sapiensP5A-2E1 LCDR1(1)..(6) 259Asn Ile Gly Ser Lys Ser1
52604PRTHomo sapiensP5A-2E1 LCDR2(1)..(3)misc_feature(4)..(4)Xaa is
absent. 260Tyr Asp Ser Xaa126111PRTHomo sapiensP5A-2E1
LCDR3(1)..(11) 261Gln Val Trp Asp Ser Ser Ser Asp His Val Val1 5
10262119PRTHomo sapiensP5A-2E1 VH(1)..(119) 262Glu Val Gln Leu Val
Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu1 5 10 15Ser Leu Lys Ile
Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30Trp Ile Gly
Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45Gly Ile
Ile Tyr Pro Gly Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe 50 55 60Gln
Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr65 70 75
80Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys
85 90 95Ala Gln Thr Ser Val Thr Arg Asn Trp Phe Asp Pro Trp Gly Gln
Gly 100 105 110Thr Leu Val Thr Val Ser Ser 115263108PRTHomo
sapiensP5A-2E1 VL(1)..(108) 263Ser Tyr Val Leu Thr Gln Pro Pro Ser
Val Ser Val Ala Pro Gly Lys1 5 10 15Thr Ala Arg Ile Thr Cys Gly Gly
Asn Asn Ile Gly Ser Lys Ser Val 20 25 30His Trp Tyr Gln Gln Lys Pro
Gly Gln Ala Pro Val Leu Val Ile Tyr 35 40 45Tyr Asp Ser Asp Arg Pro
Ser Gly Ile Pro Glu Arg Phe Ser Gly Ser 50 55 60Asn Ser Gly Asn Thr
Ala Thr Leu Thr Ile Ser Arg Val Glu Ala Gly65 70 75 80Asp Glu Ala
Asp Tyr Tyr Cys Gln Val Trp Asp Ser Ser Ser Asp His 85 90 95Val Val
Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105264357DNAHomo
sapiensP5A-2E1 VHnu(1)..(357) 264gaggtgcagc tggtgcagtc tggagcagag
gtgaaaaagc ccggggagtc tctgaagatc 60tcctgtaagg gttctggata cagctttacc
agctactgga tcggctgggt gcgccagatg 120cccgggaaag gcctggagtg
gatggggatc atctatcctg gtgactctga taccagatac 180agcccgtcct
tccaaggcca ggtcaccatc tcagccgaca agtccatcag caccgcctac
240ctgcagtgga gcagcctgaa ggcctcggac accgccatgt attactgtgc
ccagacgtca 300gtgactcgca actggttcga cccctggggc cagggaaccc
tggtcaccgt ctcctca 357265324DNAHomo sapiensP5A-2E1 VLnu(1)..(324)
265tcctatgtgc tgactcagcc accctcagtg tcagtggccc caggaaagac
ggccaggatt 60acctgtgggg gaaacaacat tggaagtaaa agtgtgcact ggtaccagca
gaagccaggc 120caggcccctg tgctggtcat ctattatgat agcgaccggc
cctcagggat ccctgagcga 180ttctctggct ccaactctgg gaacacggcc
accctgacca tcagcagggt cgaagccggg 240gatgaggccg actattactg
tcaggtgtgg gatagtagta gtgatcatgt ggtattcggc 300ggagggacca
agctgaccgt ccta 3242668PRTHomo sapiensP5A-1C8 HCDR1(1)..(8) 266Gly
Tyr Thr Phe Thr Ser Tyr Tyr1 52678PRTHomo sapiensP5A-1C8
HCDR2(1)..(8) 267Ile Asn Pro Ser Gly Gly Ser Thr1 526822PRTHomo
sapiensP5A-1C8 HCDR3(1)..(22) 268Ala Arg Ser Ala Arg Asp Tyr Tyr
Asp Ser Ser Gly Tyr Tyr Tyr Arg1 5 10 15Ala Glu Tyr Phe Gln His
202696PRTHomo sapiensP5A-1C8 LCDR1(1)..(6) 269Gln Asp Ile Ser Asn
Tyr1 52704PRTHomo sapiensP5A-1C8
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 270Asp Ala Ser
Xaa127110PRTHomo sapiensP5A-1C8 LCDR3(1)..(10) 271Gln Gln Tyr Asp
Asn Leu Pro Ser Ile Thr1 5 10272129PRTHomo sapiensP5A-1C8
VH(1)..(129) 272Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
Pro Gly Ala1 5 10 15Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr
Phe Thr Ser Tyr 20 25 30Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln
Gly Leu Glu Trp Met 35 40 45Gly Ile Ile Asn Pro Ser Gly Gly Ser Thr
Ser Tyr Ala Gln Lys Phe 50 55 60Gln Gly Arg Val Thr Met Thr Arg Asp
Thr Ser Thr Ser Thr Val Tyr65 70 75 80Met Glu Leu Ser Ser Leu Arg
Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Ser Ala Arg Asp
Tyr Tyr Asp Ser Ser Gly Tyr Tyr Tyr Arg 100 105 110Ala Glu Tyr Phe
Gln His Trp Gly Gln Gly Thr Leu Val Thr Val Ser 115 120
125Ser273108PRTHomo sapiensmisc_feature(1)..(108)P5A-1C8 VL 273Asp
Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10
15Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp Ile Ser Asn Tyr
20 25 30Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu
Ile 35 40 45Tyr Asp Ala Ser Asn Leu Glu Thr Gly Val Pro Ser Arg Phe
Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Phe Thr Ile Ser Ser
Leu Gln Pro65 70 75 80Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Gln Tyr
Asp Asn Leu Pro Ser 85 90 95Ile Thr Phe Gly Gln Gly Thr Arg Leu Glu
Ile Lys 100 105274387DNAHomo sapiensP5A-1C8
VHnu(1)..(387) 274caggtgcagc tggtgcagtc tggggctgag gtgaagaagc
ctggggcctc agtgaaggtt 60tcctgcaagg catctggata caccttcacc agctactata
tgcactgggt gcgacaggcc 120cctggacaag ggcttgagtg gatgggaata
atcaacccta gtggtggtag cacaagctac 180gcacagaagt tccagggcag
agtcaccatg accagggaca cgtccacgag cacagtctac 240atggagctga
gcagcctgag atctgaggac acggccgtgt attactgtgc gaggtcggcc
300cgggattact atgatagtag tggttattac taccgcgctg aatacttcca
gcactggggc 360cagggcaccc tggtcaccgt ctcctca 387275324DNAHomo
sapiensP5A-1C8 VLnu(1)..(324) 275gacatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc aggcgagtca ggacattagc
aactatttaa attggtatca gcagaaacca 120gggaaagccc ctaagctcct
gatctacgat gcatccaatt tggaaacagg ggtcccatca 180aggttcagtg
gaagtggatc tgggacagat tttactttca ccatcagcag cctgcagcct
240gaagatattg caacatatta ctgtcaacag tatgataatc tcccctctat
caccttcggc 300caagggacac gactggagat taaa 3242768PRTHomo
sapiensP1A-1C10 HCDR1(1)..(8) 276Gly Gly Thr Ser Ser Phe Tyr Asp1
52778PRTHomo sapiensP1A-1C10 HCDR2(1)..(8) 277Ile Ile Pro Arg Leu
Asp Ile Ala1 527816PRTHomo sapiensP1A-1C10 HCDR3(1)..(16) 278Ala
Arg Gly Arg Pro Gly Ser Glu Trp Ala Tyr Gly Pro Phe Asp Leu1 5 10
152796PRTHomo sapiensP1A-1C10 KCDR1(1)..(6) 279Gln Ser Ser Arg Ala
Trp1 52804PRTHomo sapiensP1A-1C10
KCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 280Lys Ala Ser
Xaa12819PRTHomo sapiensP1A-1C10 KCDR3(1)..(9) 281His Gln Tyr Asn
Ser Ser Pro Phe Thr1 5282123PRTHomo sapiensP1A-1C10 VH(1)..(123)
282Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Asn Pro Gly Ser1
5 10 15Ser Val Lys Val Ser Cys Lys Ala Gly Gly Gly Thr Ser Ser Phe
Tyr 20 25 30Asp Ile Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu
Trp Ile 35 40 45Gly Lys Ile Ile Pro Arg Leu Asp Ile Ala Asp Tyr Ala
Gln Lys Ser 50 55 60Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr
Ser Thr Val Tyr65 70 75 80Leu Glu Leu Ser Ser Leu Lys Ser Asp Asp
Thr Ala Val Tyr Phe Cys 85 90 95Ala Arg Gly Arg Pro Gly Ser Glu Trp
Ala Tyr Gly Pro Phe Asp Leu 100 105 110Trp Gly Gln Gly Thr Leu Val
Thr Val Ser Ser 115 120283107PRTHomo sapiensP1A-1C10 VL(1)..(107)
283Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly1
5 10 15Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ser Arg Ala
Trp 20 25 30Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu
Leu Ile 35 40 45Ser Lys Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg
Phe Ser Gly 50 55 60Ser Gly Tyr Gly Thr Glu Phe Thr Leu Thr Ile Ser
Ser Leu Gln Pro65 70 75 80Asp Asp Ser Ala Thr Tyr Tyr Cys His Gln
Tyr Asn Ser Ser Pro Phe 85 90 95Thr Phe Gly Pro Gly Thr Lys Val Gln
Ile Lys 100 105284369DNAHomo sapiensP1A-1C10 VHnu(1)..(369)
284caggtgcagc tggtgcagtc tggggctgag gtgaagaacc cggggtcctc
ggtgaaggtc 60tcctgtaagg ctggtggagg cacctccagt ttctatgata tcaactgggt
gcgacaggcc 120cctggacaag ggcttgagtg gataggaaaa atcatcccta
ggcttgatat agcagactac 180gcacagaagt cccagggcag agtcacgatt
accgcggaca aatccacgag tacagtatac 240ttggaattga gcagcctgaa
gtcagacgac acggccgtgt atttctgtgc gagaggtcgg 300ccgggttcgg
agtgggcgta tggcccattt gacctctggg gccagggaac cctggtcacc 360gtctcctca
369285321DNAHomo sapiensP1A-1C10 Vlnu(1)..(321) 285gacatccaga
tgacccagtc tccttccacc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc
gggccagtca gagttctagg gcctggttgg cctggtatca gcagaaacca
120gggaaagccc ctaaactcct gatctctaag gcgtctagtt tagaaagtgg
ggtcccatca 180aggttcagcg gcagtggata tgggacagaa ttcactctca
ccatcagcag cctgcagcct 240gatgattctg caacttatta ctgccaccag
tataacagta gcccattcac tttcggccct 300gggaccaaag tgcagatcaa a
3212868PRTHomo sapiensP4A-1H6 HCDR1(1)..(8) 286Gly Phe Thr Phe Ser
Ser Tyr Gly1 52878PRTHomo sapiensP4A-1H6 HCDR2(1)..(8) 287Ile Ser
Asp Asp Gly Ser Asn Gln1 528821PRTHomo sapiensP4A-1H6
HCDR3(1)..(21) 288Ala Lys Arg Gly Gly Tyr Cys Ser Thr Thr Ser Cys
Leu Leu Arg Trp1 5 10 15Val Tyr Phe Asp Phe 202896PRTHomo
sapiensP4A-1H6 LCDR1(1)..(6) 289Gln Ser Ile Ser Ser Tyr1
52904PRTHomo sapiensP4A-1H6 LCDR2(1)..(3)misc_feature(4)..(4)Xaa is
absent 290Ala Ala Ser Xaa12918PRTHomo sapiensP4A-1H6 LCDR3(1)..(8)
291Gln Gln Ser Tyr Asn Thr Pro Thr1 5292128PRTHomo sapiensP4A-1H6
VH(1)..(128) 292Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ser Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Asp Asp Gly Ser Asn Gln
Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg
Val Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Arg Gly Gly Tyr
Cys Ser Thr Thr Ser Cys Leu Leu Arg Trp 100 105 110Val Tyr Phe Asp
Phe Trp Gly Gln Gly Thr Leu Ala Thr Val Ser Ser 115 120
125293106PRTHomo sapiensP4A-1H6 VL(1)..(106) 293Asp Ile Gln Met Thr
Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr
Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Tyr 20 25 30Leu His Trp
Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile 35 40 45Tyr Ala
Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser
Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75
80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ser Tyr Asn Thr Pro Thr
85 90 95Phe Gly Pro Gly Thr Lys Val Asp Ile Lys 100
105294384DNAHomo sapiensP4A-1H6 VHnu(1)..(384) 294caggtgcagc
tggtggagtc tgggggaggc gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag
cctctggatt caccttcagt agctatggca tgcactgggt ccgccagtct
120ccaggcaagg ggctggagtg ggtggcagtt atatcagatg atggaagtaa
tcaatactat 180gcagactccg tgaagggccg attcaccatc tccagagaca
attccaagaa cacgctgtat 240ctgcaaatga acagcctgag agttgaggac
acggctgtgt attactgtgc gaaaaggggc 300ggatattgta gtactaccag
ctgcctcctt aggtgggtct actttgactt ctggggccag 360ggaaccctgg
ccaccgtctc ctca 384295318DNAHomo sapiensP4A-1H6 Vlnu(1)..(318)
295gacatccaga tgacccagtc tccatcctcc ctgtctgcat ctgtaggaga
cagagtcacc 60atcacttgcc gggcaagtca gagcattagc agctatttac attggtatca
gcaaaaacca 120gggaaagccc ctaacctcct gatctatgct gcatccagtt
tgcaaagtgg ggtcccatca 180aggttcagtg gcagtggatc tgggacagat
ttcactctca ccatcagcag tctgcaacct 240gaagactttg caacttacta
ctgtcaacag agttacaata cccctacttt cggccctggg 300accaaagtgg atatcaaa
3182968PRTHomo sapiensP4B-1F4 HCDR1(1)..(8) 296Gly Phe Thr Phe Ser
Ser Tyr Gly1 52978PRTHomo sapiensP4B-1F4 HCDR2(1)..(8) 297Ile Ser
Tyr Asp Gly Ser Asn Lys1 529822PRTHomo sapiensP4B-1F4
HCDR3(1)..(22) 298Ala Lys Gly Pro Arg Tyr Ser Ser Ser Trp Tyr Ile
Ser Leu Tyr Tyr1 5 10 15Tyr Tyr Gly Met Asp Val 2029911PRTHomo
sapiensP4B-1F4 LCDR1(1)..(11) 299Gln Ser Leu Val Tyr Ser Asp Gly
Asn Thr Tyr1 5 103004PRTHomo sapiensP4B-1F4
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 300Lys Val Ser
Xaa130110PRTHomo sapiensP4B-1F4 LCDR3(1)..(10) 301Met Gln Ala Thr
His Trp Pro Leu Tyr Thr1 5 10302129PRTHomo sapiensP4B-1F4
VH(1)..(129) 302Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys
Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Ile Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Lys Gly Pro Arg Tyr
Ser Ser Ser Trp Tyr Ile Ser Leu Tyr Tyr 100 105 110Tyr Tyr Gly Met
Asp Val Trp Gly Gln Gly Thr Thr Val Thr Val Ser 115 120
125Ser303113PRTHomo sapiensP4B-1F4 VL(1)..(113) 303Asp Val Val Met
Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly1 5 10 15Gln Pro Ala
Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val Tyr Ser 20 25 30Asp Gly
Asn Thr Tyr Leu Asn Trp Phe Gln Gln Arg Pro Gly Gln Ser 35 40 45Pro
Arg Arg Leu Ile Tyr Lys Val Ser Asn Arg Asp Ser Gly Val Pro 50 55
60Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile65
70 75 80Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln
Ala 85 90 95Thr His Trp Pro Leu Tyr Thr Phe Gly Gln Gly Thr Lys Leu
Glu Ile 100 105 110Lys304387DNAHomo sapiensP4B-1F4 VHnu(1)..(387)
304caggtgcagc tggtggagtc tgggggaggc gtggtccagc ctgggaggtc
cctgagactc 60tcctgtgcag cctctggatt caccttcagt agctatggca tgcactgggt
ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt atatcatatg
atggaagtaa taaatactat 180gcagactccg tgaagggccg attcaccatc
tccagagaca attccaagaa cacgctgtat 240ctgcaaatca acagcctgag
agctgaggac acggctgtgt attactgtgc gaaagggcct 300cggtatagca
gcagctggta cataagcctt tactactact acggtatgga cgtctggggc
360caagggacca cggtcaccgt ctcctca 387305339DNAHomo sapiensP4B-1F4
Vlnu(1)..(339) 305gatgttgtga tgactcagtc tccactctcc ctgcccgtca
cccttggaca gccggcctcc 60atctcctgca ggtctagtca aagcctcgta tacagtgatg
gaaacaccta cttgaattgg 120tttcagcaga ggccaggcca atctccaagg
cgcctaattt ataaggtttc taaccgggac 180tctggggtcc cagacagatt
cagcggcagt gggtcaggca ctgatttcac actgaaaatc 240agcagggtgg
aggctgagga tgttggggtt tattactgca tgcaagctac acactggccc
300ctgtacactt ttggccaggg gaccaagctg gagatcaaa 3393068PRTHomo
sapiensP5A-1B6 HCDR1(1)..(8) 306Gly Phe Thr Phe Ser Ser Tyr Ala1
53078PRTHomo sapiensP5A-1B6 HCDR2(1)..(8) 307Ile Ser Tyr Asp Gly
Ser Asn Lys1 530820PRTHomo sapiensP5A-1B6 HCDR3(1)..(20) 308Ala Arg
Asp Gly Gln Ala Ile Thr Met Val Gln Gly Val Ile Gly Pro1 5 10 15Pro
Phe Asp Tyr 203096PRTHomo sapiensP5A-1B6 LCDR1(1)..(6) 309Gln Asp
Ile Ser Asn Tyr1 53104PRTHomo sapiensP5A-1B6
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 310Asp Ala Ser
Xaa13119PRTHomo sapiensP5A-1B6 LCDR3(1)..(9) 311Gln Gln Tyr Asp Asn
Leu Pro Tyr Thr1 5312127PRTHomo sapiensP5A-1B6 VH(1)..(127) 312Gln
Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln Pro Gly Arg1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
20 25 30Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45Ala Val Ile Ser Tyr Asp Gly Ser Asn Lys Tyr Tyr Ala Asp
Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn
Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr
Ala Val Tyr Tyr Cys 85 90 95Ala Arg Asp Gly Gln Ala Ile Thr Met Val
Gln Gly Val Ile Gly Pro 100 105 110Pro Phe Asp Tyr Trp Gly Gln Gly
Thr Leu Val Thr Val Ser Ser 115 120 125313107PRTHomo sapiensP5A-1B6
VL(1)..(107) 313Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala
Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp
Ile Ser Asn Tyr 20 25 30Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala
Pro Lys Leu Leu Ile 35 40 45Tyr Asp Ala Ser Asn Leu Glu Thr Gly Val
Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr Asp Phe Thr Phe
Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Ile Ala Thr Tyr Tyr
Cys Gln Gln Tyr Asp Asn Leu Pro Tyr 85 90 95Thr Phe Gly Gln Gly Thr
Lys Leu Glu Ile Lys 100 105314381DNAHomo sapiensP5A-1B6
VHnu(1)..(381) 314caggtgcagc tggtggagtc tgggggaggc gtggtccagc
ctgggaggtc cctgagactc 60tcctgtgcag cctctggatt caccttcagt agctatgcta
tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg ggtggcagtt
atatcatatg atggaagtaa taaatactac 180gcagactccg tgaagggccg
attcaccatc tccagagaca attccaagaa cacgctgtat 240ctgcaaatga
acagcctgag agctgaggac acggctgtgt attactgtgc gagagatgga
300caggctatta ctatggttca gggagttatc ggcccaccct ttgactactg
gggccaggga 360accctggtca ccgtctcctc a 381315321DNAHomo
sapiensP5A-1B6 Vlnu(1)..(321) 315gacatccaga tgacccagtc tccatcctcc
ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc aggcgagtca ggacattagc
aactatttaa attggtatca gcagaaacca 120gggaaagccc ctaagctcct
gatctacgat gcatccaatt tggaaacagg ggtcccatca 180aggttcagtg
gaagtggatc tgggacagat tttactttca ccatcagcag cctgcagcct
240gaagatattg caacatatta ctgtcaacag tatgataatc tcccgtacac
ttttggccag 300gggaccaagc tggagatcaa a 3213168PRTHomo sapiensP5A-1B8
HCDR1(1)..(8) 316Gly Phe Thr Val Ser Ser Asn Tyr1 53177PRTHomo
sapiensP5A-1B8 HCDR2(1)..(7) 317Ile Tyr Pro Gly Gly Ser Thr1
53189PRTHomo sapiensP5A-1B8 HCDR3(1)..(9) 318Ala Arg Glu Thr Leu
Ala Phe Asp Tyr1 53196PRTHomo sapiensP5A-1B8 LCDR1(1)..(6) 319Gln
Gly Ile Ser Ser Tyr1 53204PRTHomo sapiensP5A-1B8
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 320Ala Ala Ser
Xaa13219PRTHomo sapiensP5A-1B8 LCDR3(1)..(9) 321Gln Gln Leu Asn Ser
Tyr Pro Pro Ala1 5322115PRTHomo sapiensP5A-1B8 VH(1)..(115) 322Glu
Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Ser Asn
20 25 30Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45Ser Val Ile Tyr Pro Gly Gly Ser Thr Phe Tyr Ala Asp Ser
Val Lys 50 55 60Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr Leu65 70 75 80Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr Cys Ala 85 90 95Arg Glu Thr Leu Ala Phe Asp Tyr Trp Gly
Gln Gly Thr Leu Val Thr 100 105 110Val Ser Ser 115323107PRTHomo
sapiensP5A-1B8 VL(1)..(107) 323Asp Ile Gln Leu Thr Gln Ser Pro Ser
Phe Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val Thr Ile Thr Cys Arg
Ala Ser Gln Gly Ile Ser Ser Tyr 20 25 30Leu Ala Trp Tyr Gln Gln Lys
Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr Ala Ala Ser Thr Leu
Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60Ser Gly Ser Gly Thr
Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65 70 75 80Glu Asp Phe
Ala Thr Tyr Tyr Cys Gln Gln Leu Asn Ser Tyr Pro Pro 85 90 95Ala Phe
Gly Gly Gly Thr Lys Val Glu Ile Lys 100 105324345DNAHomo
sapiensP5A-1B8 VHnu(1)..(345) 324gaggtgcagc tggtggagtc tggaggaggc
ttgatccagc ctggggggtc cctgagactc 60tcctgtgcag cctctgggtt caccgtcagt
agcaactaca tgagctgggt ccgccaggct 120ccagggaagg ggctggagtg
ggtctcagtt atttatcccg gtggtagcac attctacgca 180gactccgtga
agggccgatt caccatctcc agagacaatt ccaagaacac cctgtatctt
240caaatgaaca gcctgagagc cgaggacacg gccgtgtatt actgtgcgag
agagacccta 300gcctttgact actggggcca gggaaccctg gtcaccgtct cctca
345325321DNAHomo sapiensP5A-1B8 Vlnu(1)..(321) 325gacatccagt
tgacccagtc tccatccttc ctgtctgcat ctgtaggaga cagagtcacc 60atcacttgcc
gggccagtca gggcattagc agttatttag cctggtatca gcaaaaacca
120gggaaagccc ctaagctcct gatctatgct
gcatccactt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc
tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg
caacttatta ctgtcaacag cttaatagtt accctccagc tttcggcgga
300gggaccaagg tggagatcaa a 3213268PRTHomo sapiensP5A-1B9
HCDR1(1)..(8) 326Gly Gly Ser Ile Ser Ser Tyr Tyr1 53277PRTHomo
sapiensP5A-1B9 HCDR2(1)..(7) 327Ile Ser Tyr Ser Gly Ser Thr1
532822PRTHomo sapiensP5A-1B9 HCDR3(1)..(22) 328Ala Ser Asn Gly Gln
Tyr Tyr Asp Ile Leu Thr Gly Gln Pro Pro Asp1 5 10 15Tyr Trp Tyr Phe
Asp Leu 2032912PRTHomo sapiensP5A-1B9 LCDR1(1)..(12) 329Gln Ser Val
Leu Tyr Ser Ser Asn Asn Lys Asn Tyr1 5 103304PRTHomo sapiensP5A-1B9
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 330Trp Ala Ser
Xaa13319PRTHomo sapiensP5A-1B9 LCDR3(1)..(9) 331Gln Gln Tyr Tyr Ser
Thr Pro Leu Thr1 5332128PRTHomo sapiensP5A-1B9 VH(1)..(128) 332Gln
Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu1 5 10
15Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Ser Ser Tyr
20 25 30Tyr Trp Ser Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Trp
Ile 35 40 45Gly Tyr Ile Ser Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser
Leu Lys 50 55 60Ser Arg Val Thr Ile Ser Leu Asp Thr Ser Lys Asn Gln
Phe Ser Leu65 70 75 80Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala
Val Tyr Tyr Cys Ala 85 90 95Ser Asn Gly Gln Tyr Tyr Asp Ile Leu Thr
Gly Gln Pro Pro Asp Tyr 100 105 110Trp Tyr Phe Asp Leu Trp Gly Arg
Gly Thr Leu Val Thr Val Ser Ser 115 120 125333113PRTHomo
sapiensP5A-1B9 VL(1)..(113) 333Asp Ile Val Met Thr Gln Ser Pro Asp
Ser Leu Ala Val Ser Leu Gly1 5 10 15Glu Arg Ala Thr Ile Asn Cys Lys
Ser Ser Gln Ser Val Leu Tyr Ser 20 25 30Ser Asn Asn Lys Asn Tyr Leu
Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45Pro Pro Lys Leu Leu Ile
Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55 60Pro Asp Arg Phe Ser
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr65 70 75 80Ile Ser Ser
Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Gln 85 90 95Tyr Tyr
Ser Thr Pro Leu Thr Phe Gly Gly Gly Thr Lys Val Glu Ile 100 105
110Lys334384DNAHomo sapiensP5A-1B9 VHnu(1)..(384) 334caggtgcagc
tgcaggagtc gggcccagga ctggtgaagc cttcggagac cctgtccctc 60acctgcactg
tctctggtgg ctccatcagt agttactact ggagctggat ccggcagccc
120ccagggaagg gactggagtg gattgggtat atctcttaca gtgggagcac
caactacaac 180ccctccctca agagtcgagt caccatatca ctagacacgt
ccaagaacca gttctccctg 240aagctgagct ctgtgaccgc tgcggacacg
gccgtgtatt actgtgcgag caacggccag 300tattacgata ttttgactgg
tcaacctcct gactactggt acttcgatct ctggggccgt 360ggcaccctgg
tcactgtctc ctca 384335339DNAHomo sapiensP5A-1B9 Vlnu(1)..(339)
335gacatcgtga tgacccagtc tccagactcc ctggctgtgt ctctgggcga
gagggccacc 60atcaactgca agtccagcca gagtgtttta tacagctcca acaataagaa
ctacttagct 120tggtaccagc agaaaccagg acagcctcct aagctgctca
tttactgggc atctacccgg 180gaatccgggg tccctgaccg attcagtggc
agcgggtctg ggacagattt cactctcacc 240atcagcagcc tgcaggctga
agatgtggca gtttattact gtcagcaata ttatagtact 300ccgctcactt
tcggcggagg gaccaaggtg gagatcaaa 3393368PRTHomo sapiensP5A-1D1
HCDR1(1)..(8) 336Gly Leu Thr Val Ser Ser Asn Tyr1 53377PRTHomo
sapiensP5A-1D1 HCDR2(1)..(7) 337Ile Tyr Ser Gly Gly Ser Thr1
533811PRTHomo sapiensP5A-1D1 HCDR3(1)..(11) 338Ala Arg Asp Leu Tyr
Tyr Tyr Gly Met Asp Val1 5 103396PRTHomo sapiensP5A-1D1
LCDR1(1)..(6) 339Gln Gly Ile Ser Ser Tyr1 53404PRTHomo
sapiensP5A-1D1 LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent.
340Ala Ala Ser Xaa13418PRTHomo sapiensP5A-1D1 LCDR3(1)..(8) 341Gln
Gln Leu Asn Ser Tyr Pro Thr1 5342117PRTHomo sapiensP5A-1D1
VH(1)..(117) 342Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln
Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Leu Thr
Val Ser Ser Asn 20 25 30Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr
Tyr Ala Asp Ser Val Lys 50 55 60Gly Arg Phe Thr Ile Ser Arg Asp Asn
Ser Lys Asn Thr Leu Tyr Leu65 70 75 80Gln Met Asn Ser Leu Arg Ala
Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95Arg Asp Leu Tyr Tyr Tyr
Gly Met Asp Val Trp Gly Gln Gly Thr Thr 100 105 110Val Thr Val Ser
Thr 115343106PRTHomo sapiensP5A-1D1 VL(1)..(106) 343Asp Ile Gln Leu
Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr 20 25 30Leu Ala
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr
Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn Ser Tyr Pro
Thr 85 90 95Phe Gly Gln Gly Thr Arg Leu Glu Ile Lys 100
105344351DNAHomo sapiensP5A-1D1 VHnu(1)..(351) 344gaggtgcagc
tggtggagtc tggaggaggc ttgatccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctgggct caccgtcagt agcaactaca tgagctgggt ccgccaggct
120ccagggaagg ggctggagtg ggtctcagtt atttatagcg gtggtagcac
atactacgca 180gactccgtga agggccgatt caccatctcc agagacaatt
ccaagaacac gctgtatctt 240caaatgaaca gcctgagagc cgaggacacg
gccgtgtatt actgtgcgag agatttgtac 300tactacggta tggacgtctg
gggccaaggg accacggtca ccgtctccac a 351345318DNAHomo sapiensP5A-1D1
Vlnu(1)..(318) 345gacatccagt tgacccagtc tccatccttc ctgtctgcat
ctgtaggaga cagagtcacc 60atcacttgcc gggccagtca gggcattagc agttatttag
cctggtatca gcaaaaacca 120gggaaagccc ctaagctcct gatctatgct
gcatccactt tgcaaagtgg ggtcccatca 180aggttcagcg gcagtggatc
tgggacagat ttcactctca ccatcagcag cctgcagcct 240gaagattttg
caacttatta ctgtcaacag cttaatagtt accctacctt cggccaaggg
300acacgactgg agattaaa 3183468PRTHomo sapiensP5A-1D10 HCDR1(1)..(8)
346Gln Phe Thr Phe Ser Asp Tyr Ser1 53478PRTHomo sapiensP5A-1D10
HCDR2(1)..(8) 347Ile Ser Gln Ser Gly Ser Thr Ile1 534821PRTHomo
sapiensP5A-1D10 HCDR3(1)..(21) 348Ala Arg Gly Val Ser Pro Ser Tyr
Val Trp Gly Ser Tyr Arg Ser Leu1 5 10 15Tyr His Phe Asp Tyr
203499PRTHomo sapiensP5A-1D10 LCDR1(1)..(9) 349Ser Ser Asp Val Gly
Gly Tyr Asn Tyr1 53504PRTHomo sapiensP5A-1D10
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 350Asp Val Ser
Xaa135111PRTHomo sapiensP5A-1D10 LCDR3(1)..(11) 351Ser Ser Phe Thr
Ser Ser Thr Thr Val Val Val1 5 10352128PRTHomo sapiensP5A-1D10
VH(1)..(128) 352Gln Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Lys
Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gln Phe Thr
Phe Ser Asp Tyr 20 25 30Ser Met Thr Trp Ile Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ser Tyr Ile Ser Gln Ser Gly Ser Thr Ile
Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ala Lys Asn Ser Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Gly Val Ser Pro
Ser Tyr Val Trp Gly Ser Tyr Arg Ser Leu 100 105 110Tyr His Phe Asp
Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120
125353111PRTHomo sapiensP5A-1D10 VL(1)..(111) 353Gln Ser Ala Leu
Thr Gln Pro Ala Ser Val Ser Gly Ser Pro Gly Gln1 5 10 15Ser Ile Thr
Ile Ser Cys Thr Gly Thr Ser Ser Asp Val Gly Gly Tyr 20 25 30Asn Tyr
Val Ser Trp Tyr Gln Gln His Pro Gly Lys Ala Pro Lys Leu 35 40 45Met
Ile Tyr Asp Val Ser Asn Arg Pro Ser Gly Val Ser Asn Arg Phe 50 55
60Ser Ala Ser Lys Ser Gly Asn Thr Ala Ser Leu Thr Ile Ser Gly Leu65
70 75 80Gln Ala Glu Asp Glu Ala Asp Tyr Tyr Cys Ser Ser Phe Thr Ser
Ser 85 90 95Thr Thr Val Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val
Leu 100 105 110354384DNAHomo sapiensP5A-1D10 VHnu(1)..(384)
354caggtgcagc tggtggagtc tgggggaggc ttggtcaagc ctggagggtc
cctgagactc 60tcctgtgcag cctctcaatt caccttcagt gactactcca tgacctggat
ccgccaggct 120ccagggaagg ggctggagtg ggtttcatac attagtcaaa
gtggtagtac catatactac 180gcagactctg tgaagggccg attcaccatc
tccagggaca acgccaagaa ctcactgtat 240ctgcaaatga acagcctgag
agccgaggac acggccgtgt attactgtgc gagaggtgtc 300agcccatcct
acgtttgggg gagttatcgt tccttgtacc actttgacta ctggggccag
360ggaaccctgg tcaccgtctc ctca 384355333DNAHomo sapiensP5A-1D10
Vlnu(1)..(333) 355cagtctgccc tgactcagcc tgcctccgtg tctgggtctc
ctggacagtc gatcaccatc 60tcctgcactg gaaccagcag tgacgttggt ggttataact
atgtctcctg gtaccaacaa 120cacccaggca aagcccccaa actcatgatt
tatgatgtca gtaatcggcc ctcaggggtt 180tctaatcgct tctctgcctc
caagtctggc aacacggcct ccctgaccat ctctgggctc 240caggctgagg
acgaggctga ttattactgc agctcattta caagcagcac cactgtcgtg
300gtattcggcg gagggaccaa gctgaccgtc cta 3333568PRTHomo
sapiensP5A-2D11 HCDR1(1)..(8) 356Gly Tyr Ser Phe Thr Ser Tyr Trp1
53578PRTHomo sapiensP5A-2D11 HCDR2(1)..(8) 357Ile Tyr Pro Gly Asp
Ser Asp Thr1 535813PRTHomo sapiensP5A-2D11 HCDR3(1)..(13) 358Ala
Arg Arg Asp Ser Thr Tyr Gly Gly Asn Thr Asp Tyr1 5 103598PRTHomo
sapiensP5A-2D11 LCDR1(1)..(8) 359Ser Ser Asn Ile Gly Ser Asn Thr1
53604PRTHomo sapiensP5A-2D11 LCDR2(1)..(3)misc_feature(4)..(4)Xaa
is absent. 360Ser Asn Asn Xaa136111PRTHomo sapiensP5A-2D11
LCDR3(1)..(11) 361Ala Ala Trp Asp Asp Ser Leu Asn Gly Val Val1 5
10362120PRTHomo sapiensP5A-2D11 VH(1)..(120) 362Glu Val Gln Leu Val
Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Glu1 5 10 15Ser Leu Lys Ile
Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30Trp Ile Gly
Trp Val Arg Gln Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45Gly Ile
Ile Tyr Pro Gly Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe 50 55 60Gln
Gly Gln Val Thr Ile Ser Ala Asp Lys Ser Ile Ser Thr Ala Tyr65 70 75
80Leu Gln Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys
85 90 95Ala Arg Arg Asp Ser Thr Tyr Gly Gly Asn Thr Asp Tyr Trp Gly
Gln 100 105 110Gly Thr Leu Val Thr Val Ser Ser 115 120363110PRTHomo
sapiensP5A-2D11 VL(1)..(110) 363Gln Ser Val Leu Thr Gln Pro Pro Ser
Ala Ser Gly Thr Pro Gly Gln1 5 10 15Arg Val Thr Ile Ser Cys Ser Gly
Ser Ser Ser Asn Ile Gly Ser Asn 20 25 30Thr Val Asn Trp Tyr Gln Gln
Leu Pro Gly Thr Ala Pro Lys Leu Leu 35 40 45Ile Tyr Ser Asn Asn Gln
Arg Pro Ser Gly Val Pro Asp Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly
Thr Ser Ala Ser Leu Ala Ile Ser Gly Leu Gln65 70 75 80Ser Glu Asp
Glu Ala Asp Tyr Tyr Cys Ala Ala Trp Asp Asp Ser Leu 85 90 95Asn Gly
Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105
110364360DNAHomo sapiensP5A-2D11 VHnu(1)..(360) 364gaggtgcagc
tggtgcagtc tggagcagag gtgaaaaagc ccggggagtc tctgaagatc 60tcctgtaagg
gttctggata cagctttacc agctactgga tcggctgggt gcgccagatg
120cccgggaaag gcctggagtg gatggggatc atctatcctg gtgactctga
taccagatac 180agcccgtcct tccaaggcca ggtcaccatc tcagccgaca
agtccatcag caccgcctac 240ctgcagtgga gcagcctgaa ggcctcggac
accgccatgt attactgtgc gagacgggat 300tcgacctacg gtggtaacac
tgactactgg ggccagggaa ccctggtcac cgtctcctca 360365330DNAHomo
sapiensP5A-2D11 Vlnu(1)..(330) 365cagtctgtgc tgactcagcc accctcagcg
tctgggaccc ccgggcagag ggtcaccatc 60tcttgttctg gaagcagctc caacatcgga
agtaatactg taaactggta ccagcagctc 120ccaggaacgg cccccaaact
cctcatctat agtaataatc agcggccctc aggggtccct 180gaccgattct
ctggctccaa gtctggcacc tcagcctccc tggccatcag tgggctccag
240tctgaggatg aggctgatta ttactgtgca gcatgggatg acagcctgaa
tggtgtggta 300ttcggcggag ggaccaagct gaccgtccta 3303668PRTHomo
sapiensP5A-2G9 HCDR1(1)..(8) 366Gly Phe Thr Phe Ser Ser Tyr Gly1
53678PRTHomo sapiensP5A-2G9 HCDR2(1)..(8) 367Ile Trp Tyr Asp Gly
Ser Asn Lys1 536812PRTHomo sapiensP5A-2G9 HCDR3(1)..(12) 368Ala Arg
Trp Phe His Thr Gly Gly Tyr Phe Asp Tyr1 5 103699PRTHomo
sapiensP5A-2G9 LCDR1(1)..(9) 369Ser Asp Ile Asn Val Ser Ser Tyr
Asn1 53707PRTHomo sapiensP5A-2G9 LCDR2(1)..(7) 370Tyr Tyr Ser Asp
Ser Asp Lys1 537110PRTHomo sapiensP5A-2G9 LCDR3(1)..(10) 371Met Ile
Trp Pro Ser Asn Ala Leu Tyr Val1 5 10372119PRTHomo sapiensP5A-2G9
VH(1)..(119) 372Gln Val Gln Leu Val Glu Ser Gly Gly Gly Val Val Gln
Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr
Phe Ser Ser Tyr 20 25 30Gly Met His Trp Val Arg Gln Ala Pro Gly Lys
Gly Leu Glu Trp Val 35 40 45Ala Val Ile Trp Tyr Asp Gly Ser Asn Lys
Tyr Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Thr Ile Ser Arg Asp
Asn Ser Lys Asn Thr Leu Tyr65 70 75 80Leu Gln Met Asn Ser Leu Arg
Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95Ala Arg Trp Phe His Thr
Gly Gly Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110Thr Leu Val Thr
Val Ser Ser 115373116PRTHomo sapiensP5A-2G9 VL(1)..(116) 373Gln Pro
Val Leu Thr Gln Pro Pro Ser Ser Ser Ala Ser Pro Gly Glu1 5 10 15Ser
Ala Arg Leu Thr Cys Thr Leu Pro Ser Asp Ile Asn Val Ser Ser 20 25
30Tyr Asn Ile Tyr Trp Tyr Gln Gln Lys Pro Gly Ser Pro Pro Arg Tyr
35 40 45Leu Leu Tyr Tyr Tyr Ser Asp Ser Asp Lys Gly Gln Gly Ser Gly
Val 50 55 60Pro Ser Arg Phe Ser Gly Ser Lys Asp Ala Ser Ala Asn Thr
Gly Ile65 70 75 80Leu Leu Ile Ser Gly Leu Gln Ser Glu Asp Glu Ala
Asp Tyr Tyr Cys 85 90 95Met Ile Trp Pro Ser Asn Ala Leu Tyr Val Phe
Gly Thr Gly Thr Lys 100 105 110Val Thr Val Leu 115374357DNAHomo
sapiensP5A-2G9 VHnu(1)..(357) 374caggtgcagc tggtggagtc tgggggaggc
gtggtccagc ctgggaggtc cctgagactc 60tcctgtgcag cgtctggatt caccttcagt
agctatggca tgcactgggt ccgccaggct 120ccaggcaagg ggctggagtg
ggtggcagtt atatggtatg atggaagtaa taaatactat 180gcagactccg
tgaagggccg attcaccatc tccagagaca attccaagaa cacgctgtat
240ctgcaaatga acagcctgag agccgaggac acggctgtgt attactgtgc
gagatggttc 300cacacggggg ggtactttga ctactggggc cagggaaccc
tggtcaccgt ctcctca 357375348DNAHomo sapiensP5A-2G9 Vlnu(1)..(348)
375cagcctgtgc tgactcagcc accttcctcc tccgcatctc ctggagaatc
cgccagactc 60acctgcacct tgcccagtga catcaatgtt agtagctaca acatatactg
gtaccagcag 120aagccaggga gccctcccag gtatctcctg tactactact
cagactcaga taagggccag 180ggctctggag tccccagccg cttctctgga
tccaaagatg cttcagccaa tacagggatt 240ttactcatct ccgggctcca
gtctgaggat gaggctgact attactgtat gatttggcca 300agcaatgctc
tttatgtctt cggaactggg accaaggtca ccgtccta 3483768PRTHomo
sapiensP5A-2H3 HCDR1(1)..(8) 376Gly Tyr Ser Phe Thr Ser Tyr Trp1
53778PRTHomo sapiensP5A-2H3 HCDR2(1)..(8) 377Ile Tyr Pro Gly Asp
Ser Asp Thr1 537813PRTHomo sapiensP5A-2H3 HCDR3(1)..(13) 378Ala Arg
Arg Asp Ser Thr Tyr Gly Gly Asn Thr Asp Tyr1 5 103798PRTHomo
sapiensP5A-2H3 LCDR1(1)..(8) 379Ser Ser Asn Ile Gly Ser Asn Thr1
53804PRTHomo sapiensP5A-2H3
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 380Ser Asn Asn
Xaa138111PRTHomo sapiensP5A-2H3 LCDR3(1)..(11) 381Ala Ala Trp Asp
Asp Ser Leu Asn Gly Val Val1 5 10382120PRTHomo sapiensP5A-2H3
VH(1)..(120) 382Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys
Pro Gly Glu1 5 10 15Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser
Phe Thr Ser Tyr 20 25 30Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys
Gly Leu Glu Trp Met 35 40 45Gly Ile Ile Tyr Pro Gly Asp Ser Asp Thr
Arg Tyr Ser Pro Ser Phe 50 55 60Gln Gly Gln Val Thr Ile Ser Ala Glu
Lys Ser Ile Ser Thr Ala Tyr65 70 75 80Leu Gln Trp Ser Ser Leu Lys
Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95Ala Arg Arg Asp Ser Thr
Tyr Gly Gly Asn Thr Asp Tyr Trp Gly Gln 100 105 110Gly Thr Leu Val
Thr Val Ser Ser 115 120383110PRTHomo sapiensP5A-2H3 VL(1)..(110)
383Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln1
5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser
Asn 20 25 30Thr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys
Leu Leu 35 40 45Ile Tyr Ser Asn Asn Gln Arg Pro Ser Gly Val Pro Asp
Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile
Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala
Ala Trp Asp Asp Ser Leu 85 90 95Asn Gly Val Val Phe Gly Gly Gly Thr
Lys Leu Thr Val Leu 100 105 110384360DNAHomo sapiensP5A-2H3
VHnu(1)..(360) 384gaggtgcagc tggtgcagtc tggagcagag gtgaaaaagc
ccggggagtc tctgaagatc 60tcctgtaagg gttctggata cagctttacc agctactgga
tcggctgggt gcgccagatg 120cccgggaaag gcctggagtg gatggggatc
atctatcctg gtgactctga taccagatac 180agcccgtcct tccaaggcca
ggtcaccatc tcagccgaga agtccatcag caccgcctac 240ctgcagtgga
gcagcctgaa ggcctcggac accgccatgt attactgtgc gagacgggat
300tcgacctacg gtggtaacac tgactactgg ggccagggaa ccctggtcac
cgtctcctca 360385330DNAHomo sapiensP5A-2H3 Vlnu(1)..(330)
385cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag
ggtcaccatc 60tcttgttctg gaagcagctc caacatcgga agtaatactg taaactggta
ccagcagctc 120ccaggaacgg cccccaaact cctcatctat agtaataatc
agcggccctc aggggtccct 180gaccgattct ctggctccaa gtctggcacc
tcagcctccc tggccatcag tgggctccag 240tctgaggatg aggctgatta
ttactgtgca gcatgggatg acagcctgaa tggtgtggta 300ttcggcggag
ggaccaagct gaccgtccta 3303868PRTHomo sapiensP5A-3A1 HCDR1(1)..(8)
386Gly Phe Thr Val Ser Ser Asn Tyr1 53877PRTHomo sapiensP5A-3A1
HCDR2(1)..(7) 387Ile Tyr Ser Gly Gly Ser Thr1 538811PRTHomo
sapiensP5A-3A1 HCDR3(1)..(11) 388Ala Arg Asp Tyr Gly Asp Phe Tyr
Phe Asp Tyr1 5 103897PRTHomo sapiensP5A-3A1 LCDR1(1)..(7) 389Gln
Ser Val Ser Ser Ser Tyr1 53904PRTHomo sapiensP5A-3A1
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 390Gly Ala Ser
Xaa13919PRTHomo sapiensP5A-3A1 LCDR3(1)..(9) 391Gln Gln Tyr Gly Ser
Ser Pro Arg Thr1 5392117PRTHomo sapiensP5A-3A1 VH(1)..(117) 392Glu
Val Gln Leu Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly1 5 10
15Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Ser Asn
20 25 30Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp
Val 35 40 45Ser Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser
Val Lys 50 55 60Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr Leu65 70 75 80Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr Cys Ala 85 90 95Arg Asp Tyr Gly Asp Phe Tyr Phe Asp Tyr
Trp Gly Gln Gly Thr Leu 100 105 110Val Thr Val Ser Ser
115393108PRTHomo sapiensP5A-3A1 VL(1)..(108) 393Glu Ile Val Leu Thr
Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly1 5 10 15Glu Arg Ala Thr
Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30Tyr Leu Ala
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45Ile Tyr
Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60Gly
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu65 70 75
80Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro
85 90 95Arg Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 100
105394351DNAHomo sapiensP5A-3A1 VHnu(1)..(351) 394gaggtgcagc
tggtggagtc tggaggaggc ttgatccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctgggtt caccgtcagt agcaactaca tgagctgggt ccgccaggct
120ccagggaagg ggctggagtg ggtctcagtt atttatagcg gtggtagcac
atactacgca 180gactccgtga agggccgatt caccatctcc agagacaatt
ccaagaacac gctgtatctt 240caaatgaaca gcctgagagc cgaggacacg
gccgtgtatt actgtgcgag agactacggt 300gacttttact ttgactactg
gggccaggga accctggtca ccgtctcctc a 351395324DNAHomo sapiensP5A-3A1
Vlnu(1)..(324) 395gaaattgtgt tgacgcagtc tccaggcacc ctgtctttgt
ctccagggga aagagccacc 60ctctcctgca gggccagtca gagtgttagc agcagctact
tagcctggta ccagcagaaa 120cctggccagg ctcccaggct cctcatctat
ggtgcatcca gcagggccac tggcatccca 180gacaggttca gtggcagtgg
gtctgggaca gacttcactc tcaccatcag cagactggag 240cctgaagatt
ttgcagtgta ttactgtcag cagtatggta gctcacctcg cacttttggc
300caggggacca agctggagat caaa 3243968PRTHomo sapiensP5A-3A6
HCDR1(1)..(8) 396Gly Phe Thr Phe Asp Asp Tyr Ala1 53978PRTHomo
sapiensP5A-3A6 HCDR2(1)..(8) 397Ile Ser Trp Asn Ser Gly Thr Ile1
539827PRTHomo sapiensP5A-3A6 HCDR3(1)..(27) 398Ala Gly Gly Gly Thr
Met Val Arg Gly Val Ile Ala Gly Gly Gly Thr1 5 10 15His Pro Val Asp
Asp Tyr Tyr Gly Met Asp Val 20 253999PRTHomo sapiensP5A-3A6
LCDR1(1)..(9) 399Ser Ser Asp Val Gly Gly Tyr Asn Tyr1 54004PRTHomo
sapiensP5A-3A6 LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent.
400Asp Val Ser Xaa140110PRTHomo sapiensP5A-3A6 LCDR3(1)..(10)
401Ser Ser Tyr Thr Ser Ser Ser Thr Val Val1 5 10402134PRTHomo
sapiensP5A-3A6 VH(1)..(134) 402Glu Val Gln Leu Val Glu Ser Gly Gly
Gly Leu Val Gln Pro Gly Arg1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Phe Asp Asp Tyr 20 25 30Ala Met His Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Gly Ile Ser Trp Asn
Ser Gly Thr Ile Gly Tyr Ala Asp Ser Val 50 55 60Lys Gly Arg Phe Ile
Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr65 70 75 80Leu Gln Met
Asn Ser Leu Arg Ala Glu Asp Thr Ala Leu Tyr Tyr Cys 85 90 95Ala Gly
Gly Gly Thr Met Val Arg Gly Val Ile Ala Gly Gly Gly Thr 100 105
110His Pro Val Asp Asp Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr
115 120 125Thr Val Thr Val Ser Ser 130403110PRTHomo sapiensP5A-3A6
VL(1)..(110) 403Gln Ser Ala Leu Thr Gln Pro Ala Ser Val Ser Gly Ser
Pro Gly Gln1 5 10 15Ser Ile Thr Ile Ser Cys Thr Gly Thr Ser Ser Asp
Val Gly Gly Tyr 20 25 30Asn Tyr Val Ser Trp Tyr Gln Gln His Pro Gly
Lys Ala Pro Lys Leu 35 40 45Met Ile Tyr Asp Val Ser Asn Arg Pro Ser
Gly Val Ser Asn Arg Phe 50 55 60Ser Gly Ser Lys Ser Gly Asn Thr Ala
Ser Leu Thr Ile Ser Gly Leu65 70 75 80Gln Ala Glu Asp Glu Ala Asp
Tyr Tyr Cys Ser Ser Tyr Thr Ser Ser 85 90 95Ser Thr Val Val Phe Gly
Gly Gly Thr Lys Leu Thr Val Leu 100 105 110404402DNAHomo
sapiensP5A-3A6 VHnu(1)..(402) 404gaagtgcagc tggtggagtc tgggggaggc
ttggtacagc ctggcaggtc cctgagactc 60tcctgtgcag cctctggatt cacctttgat
gattatgcca tgcactgggt ccggcaagct 120ccagggaagg gcctggagtg
ggtctcaggt attagttgga atagtggtac cataggctat 180gcggactctg
tgaagggccg attcatcatc tccagagaca acgccaagaa ctccctgtat
240ctgcaaatga acagtctgag agctgaggac acggccttgt attactgtgc
agggggtggt 300actatggttc ggggagttat tgccggaggg ggaactcatc
cggtggatga ctactacggt 360atggacgtct ggggccaagg gaccacggtc
accgtctcct ca 402405330DNAHomo sapiensP5A-3A6 Vlnu(1)..(330)
405cagtctgccc tgactcagcc tgcctccgtg tctgggtctc ctggacagtc
gatcaccatc 60tcctgcactg gaaccagcag tgacgttggt ggttataact atgtctcctg
gtaccaacaa 120cacccaggca aagcccccaa actcatgatt tatgatgtca
gtaatcggcc ctcaggggtt 180tctaatcgct tctctggctc caagtctggc
aacacggcct ccctgaccat ctctgggctc 240caggctgagg acgaggctga
ttattactgc agctcatata caagcagcag cactgtggta 300ttcggcggag
ggaccaagct gaccgtccta 3304068PRTHomo sapiensP5A-3B4 HCDR1(1)..(8)
406Gly Tyr Ser Phe Thr Ser Tyr Trp1 54078PRTHomo sapiensP5A-3B4
HCDR2(1)..(8) 407Ile Tyr Pro Gly Asp Ser Asp Thr1 540813PRTHomo
sapiensP5A-3B4 HCDR3(1)..(13) 408Ala Arg Arg Asp Ser Thr Tyr Gly
Gly Asn Thr Asp Tyr1 5 104098PRTHomo sapiensP5A-3B4 LCDR1(1)..(8)
409Ser Ser Asn Ile Gly Ser Asn Thr1 54104PRTHomo sapiensP5A-3B4
LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent. 410Ser Asn Asn
Xaa141111PRTHomo sapiensP5A-3B4 LCDR3(1)..(11) 411Ala Ala Trp Asp
Asp Ser Leu Asn Gly Val Val1 5 10412120PRTHomo sapiensP5A-3B4
VH(1)..(120) 412Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Glu
Pro Gly Glu1 5 10 15Ser Leu Lys Ile Ser Cys Lys Gly Ser Gly Tyr Ser
Phe Thr Ser Tyr 20 25 30Trp Ile Gly Trp Val Arg Gln Met Pro Gly Lys
Gly Leu Glu Trp Met 35 40 45Gly Ile Ile Tyr Pro Gly Asp Ser Asp Thr
Arg Tyr Ser Pro Ser Phe 50 55 60Gln Gly Gln Val Thr Ile Ser Ala Asp
Lys Ser Ile Ser Thr Ala Tyr65 70 75 80Leu Gln Trp Ser Ser Leu Lys
Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95Ala Arg Arg Asp Ser Thr
Tyr Gly Gly Asn Thr Asp Tyr Trp Gly Gln 100 105 110Gly Thr Leu Val
Thr Val Ser Ser 115 120413110PRTHomo sapiensP5A-3B4 VL(1)..(110)
413Gln Ser Val Leu Thr Gln Pro Pro Ser Ala Ser Gly Thr Pro Gly Gln1
5 10 15Arg Val Thr Ile Ser Cys Ser Gly Ser Ser Ser Asn Ile Gly Ser
Asn 20 25 30Thr Val Asn Trp Tyr Gln Gln Leu Pro Gly Thr Ala Pro Lys
Leu Leu 35 40 45Ile Tyr Ser Asn Asn Gln Arg Pro Ser Gly Val Pro Asp
Arg Phe Ser 50 55 60Gly Ser Lys Ser Gly Thr Ser Ala Ser Leu Ala Ile
Ser Gly Leu Gln65 70 75 80Ser Glu Asp Glu Ala Asp Tyr Tyr Cys Ala
Ala Trp Asp Asp Ser Leu 85 90 95Asn Gly Val Val Phe Gly Gly Gly Thr
Lys Leu Thr Val Leu 100 105 110414360DNAHomo sapiensP5A-3B4
VHnu(1)..(360) 414gaggtgcagc tggtgcagtc tggagcagag gtgaaagagc
ccggggagtc tctgaagatc 60tcctgtaagg gttctggata cagctttacc agctactgga
tcggctgggt gcgccagatg 120cccgggaaag gcctggagtg gatggggatc
atctatcctg gtgactctga taccagatac 180agcccgtcct tccaaggcca
ggtcaccatc tcagccgaca agtccatcag caccgcctac 240ctgcagtgga
gcagcctgaa ggcctcggac accgccatgt attactgtgc gagacgggat
300tcgacctacg gtggtaacac tgactactgg ggccagggaa ccctggtcac
cgtctcctca 360415330DNAHomo sapiensP5A-3B4 Vlnu(1)..(330)
415cagtctgtgc tgactcagcc accctcagcg tctgggaccc ccgggcagag
ggtcaccatc 60tcttgttctg gaagcagctc caacatcgga agtaatactg taaactggta
ccagcagctc 120ccaggaacgg cccccaaact cctcatctat agtaataatc
agcggccctc aggggtccct 180gaccgattct ctggctccaa gtctggcacc
tcagcctccc tggccatcag tgggctccag 240tctgaggatg aggctgatta
ttactgtgca gcatgggatg acagcctgaa tggtgtggta 300ttcggcggag
ggaccaagct gaccgtccta 33041610PRTHomo sapiensP5A-3C12
HCDR1(1)..(10) 416Gly Phe Ser Leu Ser Thr Ser Gly Val Gly1 5
104177PRTHomo sapiensP5A-3C12 HCDR2(1)..(7) 417Ile Tyr Trp Asp Asp
Asp Lys1 541819PRTHomo sapiensP5A-3C12 HCDR3(1)..(19) 418Ala His
Ser Leu Phe Leu Thr Val Gly Tyr Ser Ser Ser Trp Ser Pro1 5 10 15Phe
Asp Tyr41912PRTHomo sapiensP5A-3C12 LCDR1(1)..(12) 419Gln Ser Val
Leu Tyr Ser Ser Asn Asn Lys Asn Tyr1 5 104204PRTHomo
sapiensP5A-3C12 LCDR2(1)..(3)misc_feature(4)..(4)Xaa is absent.
420Trp Ala Ser Xaa14219PRTHomo sapiensP5A-3C12 LCDR3(1)..(9) 421Gln
Gln Tyr Tyr Ser Thr Pro His Thr1 5422127PRTHomo sapiensP5A-3C12
VH(1)..(127) 422Gln Ile Thr Leu Lys Glu Ser Gly Pro Thr Leu Val Lys
Pro Thr Gln1 5 10 15Thr Leu Thr Leu Thr Cys Thr Phe Ser Gly Phe Ser
Leu Ser Thr Ser 20 25 30Gly Val Gly Val Gly Trp Ile Arg Gln Pro Pro
Gly Lys Ala Leu Glu 35 40 45Trp Leu Ala Leu Ile Tyr Trp Asp Asp Asp
Lys Arg Tyr Ser Pro Ser 50 55 60Leu Lys Ser Arg Leu Thr Ile Thr Lys
Asp Thr Ser Lys Asn Gln Val65 70 75 80Val Leu Thr Met Thr Asn Met
Asp Pro Val Asp Thr Ala Thr Tyr Tyr 85 90 95Cys Ala His Ser Leu Phe
Leu Thr Val Gly Tyr Ser Ser Ser Trp Ser 100 105 110Pro Phe Asp Tyr
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 115 120
125423113PRTHomo sapiensP5A-3C12 VL(1)..(113) 423Asp Ile Val Met
Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly1 5 10 15Glu Arg Ala
Thr Ile Asn Cys Lys Ser Ser Gln Ser Val Leu Tyr Ser 20 25 30Ser Asn
Asn Lys Asn Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 35 40 45Pro
Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 50 55
60Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr65
70 75 80Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln
Gln 85 90 95Tyr Tyr Ser Thr Pro His Thr Phe Gly Gln Gly Thr Lys Leu
Glu Ile 100 105 110Lys424381DNAHomo sapiensP5A-3C12 VHnu(1)..(381)
424cagatcacct tgaaggagtc tggtcctacg ctggtgaaac ccacacagac
cctcacgctg 60acctgcacct tctctgggtt ctcactcagc actagtggag tgggtgtggg
ctggatccgt 120cagcccccag gaaaggccct ggagtggctt gcactcattt
attgggatga tgataagcgc 180tacagcccat ctctgaagag caggctcacc
atcaccaagg acacctccaa aaaccaggtg 240gtccttacaa tgaccaacat
ggaccctgtg gacacagcca catattactg tgcacacagt 300ttgtttctca
cggtagggta tagcagcagc tggtcccctt ttgactactg gggccaggga
360accctggtca ccgtctcctc a 381425339DNAHomo sapiensP5A-3C12
Vlnu(1)..(339) 425gacatcgtga tgacccagtc tccagactcc ctggctgtgt
ctctgggcga gagggccacc 60atcaactgca agtccagcca gagtgtttta tacagctcca
acaataagaa ctacttagct 120tggtaccagc agaaaccagg acagcctcct
aagctgctca tttactgggc atctacccgg 180gaatccgggg tccctgaccg
attcagtggc agcgggtctg ggacagattt cactctcacc 240atcagcagcc
tgcaggctga agatgtggca gtttattact gtcagcaata ttatagtact
300cctcacactt ttggccaggg gaccaagctg gagatcaaa 3394268PRTHomo
sapiensP22A-1D1 HCDR1(1)..(8) 426Gly Phe Thr Val Ser Ser Asn Tyr1
54277PRTHomo sapiensP22A-1D1 HCDR2(1)..(7) 427Ile Tyr Ser Gly Gly
Ser Thr1 542811PRTHomo sapiensP22A-1D1 HCDR3(1)..(11) 428Ala Arg
Asp Arg Asp Tyr Tyr Gly Met Asp Val1 5 104296PRTHomo
sapiensP22A-1D1 LCDR1(1)..(6) 429Gln Gly Ile Ser Ser Tyr1
54304PRTHomo sapiensP22A-1D1 LCDR2(1)..(3)misc_feature(4)..(4)Xaa
is absent. 430Ala Ala Ser Xaa14318PRTHomo sapiensP22A-1D1
LCDR3(1)..(8) 431Leu His Leu Asn Ser Tyr Arg Thr1 5432117PRTHomo
sapiensP22A-1D1 VH(1)..(117) 432Glu Val Gln Leu Val Glu Ser Gly Gly
Gly Leu Ile Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys Ala Ala
Ser Gly Phe Thr Val Ser Ser Asn 20 25 30Tyr Met Ser Trp Val Arg Gln
Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Val Ile Tyr Ser Gly
Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60Gly Arg Phe Thr Ile
Ser Arg Asp Asn Ser Lys Asn Thr
Leu Tyr Leu65 70 75 80Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala
Val Tyr Tyr Cys Ala 85 90 95Arg Asp Arg Asp Tyr Tyr Gly Met Asp Val
Trp Gly Gln Gly Thr Thr 100 105 110Val Thr Val Ser Ser
115433106PRTHomo sapiensP22A-1D1 VL(1)..(106) 433Asp Ile Gln Leu
Thr Gln Ser Pro Ser Phe Leu Ser Ala Ser Val Gly1 5 10 15Asp Arg Val
Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Tyr 20 25 30Leu Ala
Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45Tyr
Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55
60Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro65
70 75 80Glu Asp Phe Ala Thr Tyr Tyr Cys Leu His Leu Asn Ser Tyr Arg
Thr 85 90 95Phe Gly Leu Gly Thr Lys Val Glu Ile Lys 100
105434351DNAHomo sapiensP22A-1D1 VHnu(1)..(351) 434gaggtgcagc
tggtggagtc tggaggaggc ttgatccagc ctggggggtc cctgagactc 60tcctgtgcag
cctctgggtt caccgtcagt agcaactaca tgagctgggt ccgccaggct
120ccagggaagg ggctggagtg ggtctcagtt atttatagcg gtggtagcac
atactacgca 180gactccgtga agggccgatt caccatctcc agagacaatt
ccaagaacac gctgtatctt 240caaatgaaca gcctgagagc cgaggacacg
gccgtgtatt actgtgcgag agatcgagac 300tactacggta tggacgtctg
gggccaaggg accacggtca ccgtctcctc a 351435318DNAHomo sapiensP22A-1D1
Vlnu(1)..(318) 435gacatccagt tgacccagtc tccatccttc ctgtctgcat
ctgtaggaga cagagtcacc 60atcacttgcc gggccagtca gggcattagc agttatttag
cctggtatca gcaaaaacca 120gggaaagccc ctaagctcct gatctatgct
gcatccactt tgcaaagtgg ggtcccatca 180aggtttagcg gcagtggatc
tgggacagaa ttcactctca caatcagcag cctgcagcct 240gaagattttg
caacttatta ctgtctacac cttaatagtt acaggacgtt cggcctaggg
300accaaggtgg aaatcaaa 31843697PRTHomo sapiens 436Glu Val Gln Leu
Val Glu Ser Gly Gly Gly Leu Ile Gln Pro Gly Gly1 5 10 15Ser Leu Arg
Leu Ser Cys Ala Ala Ser Gly Phe Thr Val Ser Ser Asn 20 25 30Tyr Met
Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser
Val Ile Tyr Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 50 55
60Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu65
70 75 80Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
Ala 85 90 95Arg43797PRTHomo sapiens 437Glu Val Gln Leu Val Glu Ser
Gly Gly Gly Leu Val Gln Pro Gly Gly1 5 10 15Ser Leu Arg Leu Ser Cys
Ala Ala Ser Gly Phe Thr Val Ser Ser Asn 20 25 30Tyr Met Ser Trp Val
Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45Ser Val Ile Tyr
Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 50 55 60Gly Arg Phe
Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu65 70 75 80Gln
Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr His Cys Ala 85 90
95Arg
* * * * *
References